MST-Department of Medical Laboratory Sciences

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Transfection and Characterization of Plasmodium Knowlesi Malaria Parasites Genetically Modified to Express Host Interferon Gamma (ifn-y) in Baboon (papio anubis)
(2008-12) Munyao, J.M.
Despite efforts to control malaria. it still remains a major health problem in endemic countries. With the development of transfection technology, it is now possible to determine the structure-function relationship of vaccine candidates. In addition, transgenes relevant in malaria vaccine development can now be expressed and host parasite interface determined. This study involved characterization of genetically modified P. knowlesi malaria parasites that express host IFN-gamma, in order to develop a baboon (Papio anubis) model for transfection and analysis of host-parasite interface. One olive baboon was infected with wild-type P. knowlesi to generate parasites for transfection. At day 9 post inoculation, the baboon developed acute malaria with a parasitaemia of 7.6% and on day 10 post inoculation it was totally bled out. The blood was processed by centrifugation and 500 ml of top brown layer harvested. This layer contained 0.65 x 109 parasites enriched with mature schizonts. The parasites were resuspended in 300 ml of cytomix, mixed with 50 ~l of plasmids DNA construct. The construct contained pyrimethamine resistant form of dihydrofolate reductase thymidylate synthase (dhfr-ts) from Toxoplasma gondii, IFN-gamma gene under control of P. berghei promoters and enhancer sequences. The mixture was electroporated in a Biorad gene pulsar in a pulse of 2.5 KV, at a capacitance of 25 ~F and a resistance of 200 nfor 0.9 ms, chilled on ice for 5 minutes and injected into two olive baboons. The baboons were given pyrimethamine orally for drug selection of transfected parasites. This was done 24 hours post inoculation. Pyrimethamine resistant P. knowlesi appeared from day 10 post inoculation. On day 15 post inoculation, blood was collected from the two baboons, and subjected to plasmodipur filtration to remove leucocytes before DNA isolation. The parasites were cultured in vitro for 48 hours, supernatant harvested and analysed by sandwich Enzyme-Linked Immunosorbent Assay (ELISA) for IFN gamma. Antibody response against transfected and passaged parasites in baboon was determined by ELISA from serum collected from baboons. Molecular characterization of isolated DNA from pyrimethamine resistant parasites by peR revealed presence of selectable marker gene (dhfr-ts) and IFN-gamma gene. Supernatant of in vitro culture of transfected parasites showed release of high levels of IFN-y, suggesting presence of IFN-gamma gene in transfected parasites. Antibody response against transfected P. knowlesi and passaged transfected parasites, suggested primary response with rise in IgM production at 0.34% and 0.33% parasitaemia respectively. These results showed for the first time that it is possible to genetically modify P. knowlesi to express host IFN-y and induce immune response in baboon. These studies have developed an in vivo transfection protocol for P. knowlesi in the baboon. This opens possibilities for using the P. knowles i-baboon model in vaccine development using cutting edge technology.
Factors predisposing the community to visceral leishmiasis in Kacheliba Division of West Pokot district, Kenya
(2011-07-15) Lotukoi, Joseph A; Orinda, G. O.
Visceral Leishmaniasis (Kala-azar) is a vector borne disease caused by obligate intramacrophage protozoan parasites. It is an old, largely unknown, a forgotten and neglected disease yet is a public health problem, a debilitating disease causing an estimated 500,000 new cases each year, and a tenth of these patients will die in the predisposed areas. The actual toll death from the disease may be higher than this estimate considering the existence of its unidentified foci. Since 1993, the regions that are Kala-azar endemic have expanded significantly, accompanied by a sharp increase in the number of recorded cases. The increasing number of people infected with the disease, poses a major health challenge because it is a silent killer, invariably killing almost all untreated patients. Currently, estimates also suggest an overall prevalence of 12 million people infected with Kala-azar in an at risk population of 350 million, suggesting more than 2 million new infectious each year with the figures including only cases with the overt disease. The new trend risks causing a public health crisis in weak Africa economies like Kenya since the vaccine for the disease is non existent. In Kenya it is common in arid and semi - arid regions of North Eastern and Rift valley provinces, especially the Kacheliba Division of West Pokot District that has a prevalence of 30% amongst other areas of Turkana District, Machakos, North Eastern, Marigat and Baringo East. The study was conducted between December, 2007 and February, 2008. Descriptive cross- sectional research design was used to determine socio demographic characteristics, economic and cultural factors, health seeking behaviour, the disease morbidity and mortality, local peoples knowledge, perception and behaviour towards the existence of Kala-azar in the area. Simple random sampling technique was used to identify study subjects in the purposively selected Kacheliba Division. A sample size of 323 respondents who were household heads or adult members and the health facility workers were randomly sampled. The data collected were processed and analyzed using the Statistical Package for Social Sciences (SPSS). The chi - square test was used to compare the relationship between variables. The key factors associated with the community being predisposed to Kala-azar include: Age, gender, educational level, presence of large number of termite mounds all over the area (76.4%), low usage of bed nets ( 25%) , inaccessibility to health services and lack of proper knowledge on transmission of the disease. Also, human activities such as hunting and deforestation (53.3%), resting or sitting near termite mounds (80.2%) and dancing at night -Adong,o ( 62.5%), when the sand flies are active. There was a significant association between age ( P= 0.001, df 2, x2=14.462) and being predisposed to Kala-azar, gender ( p= 0.001, df=4, x2=61.04), educational level ( p=0.001,df=9,x2=149.55), presence of large number of termite mounds ( p=0.001,df=8,x2=39.821) and resting or sitting near termite mounds ( p=0.001,df=2,x2=17.67). The study concludes that Kala-azar is still prevalent in the area, low economic status, inaccessibility to health services, abundant presence of termite mounds that harbour sand flies and the community's different beliefs about transmission are risk factors. The study recommends the need for enhanced general health education and awareness on the transmission cycle of Kala- azar. Community participation as well as culturally appropriate behaviour change communication activities should be emphasized to enhance strategies targeting vector control. In addition, integrated disease surveillance response to be implemented to avert the disease situation.
Genetic diversity of HIV-1 Based on GAG region among antenatal clinic attendees in North- Rift valley, Kenya
(2011-08-10) Nyagaka, Benuel
HIV and AIDS remain a major health burden worldwide. Majority of HIV and AIDS impact is borne by women. Among these women, expectant mothers who become infected by HIV and AIDS will transmit the infection to their infants. This can occur during pregnancy, labour and delivery, or breastfeeding. In order to deter HIV-1 virus spread to the unborn infants intervention strategies namely Antirctroviral (ARV) drugs and vaccines must be used. However, ARVs develop resistance due to evolution of new strains of HIV virus which are drug resistant. This suggests that new intervention strategies have to be developed to deter the spread to unborn infants. One of the options is to characterize the HIV virus in order to know the virus strains circulating among the infected population. The unique characteristics in the virus can be targeted in development of new ARVs, diagnostic techniques and vaccines. The aim of this study was to determine HIV-1 gag gene subtypes circulating among antenatal clinic clients in North-Rift Valley, Kenya. A total of 129 patients were sampled and used in this study. Peripheral blood mononuclear cells (PBMCs) were separated from whole blood. Proviral DNA was isolated from PBMCs. Nested polymerase chain reaction (PCR) was carried out to amplify the viral DNA. Presequencing polymerase chain reaction (PCR) was carried out using nested gag primers. Sequencing was carried out and an automated DNA sequencer (ABI Prism 310) using Rhodamine Terminator cycle sequencing ready reaction kit. After sequencing, the generated sequences were analyzed by comparing them with available sequences at the Los Alamos HIV database using the basic Local Alignment Search Tool (BLAST) and then aligned using CLUSTAL W software. Phylogenetic trees were then constructed using the neighbor joining method. Evolutionary tree were used to monitor the extent of HIV-1 subtype variations in infected populations. On phylogenetic analysis of the 117 sequences based on partial gag gene. 81, 70%) sequences clustered with subtype Al, 13 (I1%) with subtype D, 8 (7%) with subtype C, 5 (4%)with subtype AID, 4 (3%) with subtype AIC, 3 (3%) with subtype A2, 1 (1%) with subtype A2C, and 1 (1%) with subtype G. These results suggest that HIV-1 epidemic may be evolving towards more complex subtypes through recombination due to viral mixing. Subtype surveillance of the circulating HIV-1 subtypes is therefore important in designing AIDS vaccines and determining which antigens need to be included in diagnostic testing kits for HIV-1. The HIV-1 antigens in the new strains of HIV-1 virus revealed in this study should be included in the design and development of future anti-HIV-1 vaccine, antiretroviral drugs and diagnostic techniques.
The antimicrobial susceptibility pattern of campylobacter isolates from Nairobi, Kenya
(2011-11-17) Kabiru, Nyawira Pity; Ngeranwa, J.J.N.; Orinda, G. O.; Revathi, G.
Campylobacteriosis is a group of infections caused by Gram negative rods of genus Campylobacter. It is among the most common bacterial infections of humans worldwide. Campylobacteriosis in Kenya is managed and treated presumptively according to its clinical features and manifestation. The common antimicrobial drugs used are: fluoroquinolones, macrolides and quinolones. Indiscriminate use of these drugs may lead to development of antimicrobial resistance. The main objective of this study was to investigate the antimicrobial susceptibility pattern of Campylobacter in Aga Khan University Hospital, Nairobi, Kenya. Four hundred and forty seven (447) diarrheic stool samples were obtained from in and oust-patients seeking treatment in the Hospital. For isolation of Campylobacter, stool samples were cultured onto blood-free selective Campylobacter medium. The isolation rates of bacterial pathogens were 29 (6.5%) for Campylobacter species. Other bacteria isolated were Shigella, Salmonella and Enteropathogenic Escherichia coli. The Campylobacter isolates were characterized to species and sub-species by colonial morphology, staining, motility and Analytical Profile Index (API) Campy strip tests. Out of 29 Campylobacter isolates 28 (96.6%) were C. jejuni and one (3.4%) was C. coh. The proportional distribution of C. jejuni bio-type 1 accounted for six (20.7%) while bio-type 2 accounted for 23 (79.3%). There was significant difference between isolation rates of Campylobacter across the age groups (x2 = 8.825, p = 0.032. There was no significant difference in isolation of Campylobacter between males and females (x2 = 0.534, p = 0.465). Out of the 300 samples that had invasive infection 15 (5%) were due to Campylobacter. Antimicrobial susceptibility testing was performed by disk diffusion method and by E-test system according to Clinical and Laboratory Standards Institute (CLST). The study findings showed that there was no resistance to azithromycin, chloramphenicol,, gentamicin and doxycycline. Resistance to ciprofloxacin was detected in four (13.8 %), ampicillin two (7.1 %), nalidixic acid three (10.3 %), cotrimoxazole 23 (79.3 %). Cross-resistance was detected between ciprofloxacin and nalidixic acid and between cotrimoxazole and ampicillin. Beta-lactamase production was detected in 75.5% of the isolates. The results of this study showed that the prevalence of Campylobacter in the patients from Aga Khan University Hospital was 6.5%. There was detection of resistance of the Campylobacter isolates to some antimicrobial drugs commonly used. Beta-lactamase production occurred in many isolates. It is therefore important for the treatment of Campylobacter to be instituted only after susceptibility testing has been done.
Isolation, identification and susceptibility profile of candida species to antifungal agents in pregnant women in Thika District Hospital, Kenya
(2012-04-13) Chengo, Nelson Menza; Margaret W. Muturi; M.W.Wanyoike
Vaginal candidiasis is relatively higher in pregnant women particularly inthe third trimester compared to nonpregnant women. Infection with Candida species causes choriomnionitis which is associated with pre-term delivery and death of the infant. Therefore, epidemiological monitoring of vaginal candidiasis infections is highly desirable for continuous determination of the existing causative species and the disease trends. Determination of susceptibility profile will be of importance in the appropriate selection of drugs for effective treatment of the infection in pregnant women. The objectives of the study were to determine the prevalence of vaginal candidiasis and identify the Candida species. The study also aimed at determining the susceptibility profile of the Candida species to antifungal drugs prescribed to the pregnant women with symptoms of the infection. A cross sectional study design and purposive sampling techniques were adopted in this study. Vaginal swabs were collected from the pregnant women attending the antenatal clinic of Thika District hospital between the months of June and August, 2010. The samples were gram stained and inoculated on Sabourands Dextrose Agar (SDA). Isolates from SDA were plated on CHROMagar for detection of mixed cultures. Germ tube, chlamydospore formation and sugar assimilation tests were performed for identification of isolates. Susceptibility profile was done using broth microdilution minimum inhibiting concentration method based on the approved NCCLS, 2002. The prevalence of vaginal candidiasis was 42.7%. The distribution of vaginal candidiasis had the highest incidence of 60% in women aged 26-35years followed by those aged 15-25years who were 26%. Twelve per cent (12%) of the women infected were aged 36-45 years. The women aged 46 and above years were the least infected (2%). This study showed that Candida albicans was the most common vaginal Candida species causing vaginal candidiasis in all the age groups. It was also found to be the most abundant species of all Candida species isolated. The women in their 3rd trimester had the highest infection occurrence of 68.09% followed by those in the 2nd trimester with 21.28% while those in their first trimester of pregnancy (10.63%) were least infected. Candida albieans was the most isolated species with 63.8%, followed by Candida glabrata 29.79%, Candida tropicalis 3.19%, Candida krusei 2.13% and Candida parapsilosis was the least with 1.06%. Most of the vaginal Candida species were isolated in 60% women who were 26-35 years, followed by 26% of those between 15-25 years. Only 2% of the women with 46years and above years were infected. The azole antifungal agents (fluconazole, ketoconazole, itraconazole and c1otrimazole) used showed high efficacy against all the vaginal Candida species isolated except Candida krusei. Candida krusei showed 100% resistance to fluconazole, c1otrimazole and a 50% resistance to ketoconazole. It was only susceptible (100%) to itraconazole. The other non-albieans Candida species were susceptible to the azole antifungal drugs. Candida albicans, Candida glabrata and Candida krusei showed moderate susceptibility (66.67%, 57.14% and 50%) respectively to topical nystatin. Candida parapsilosis was 100% susceptible to the drug. Nonetheless, the susceptibility of vaginal Candida species to the azole drugs and topical nystatin observed in this study supports the continued use of azole drugs but not topical nystatin for the treatment of vaginal candidiasis in the pregnant women attending the Thika District Hospital.
The bacteriologic quality of Dandora sewage treatment plant and the receiving waters of Nairobi and Athi Rivers
(2012-04-24) Musyoki, Abednego Moki
Wastewater is known to contain microbes that are deleterious to human health. Epidemiological reports show that wastewater associated disease outbreaks are common around the world. This suggests that wastewater must be processed carefully before release into natural waters and the environment. The purpose of this study was to determine the total bacterial load and microbial types in Dandora Sewage Treatment Plant (DSTP) and its handling capacity in terms of pathogen removal. In addition, the seasonal and temporal relationship of bacterial load variation was estimated. Antecedent bacterial load of Nairobi and Athi rivers was also established to reveal the microbial load emptied into Athi River by the DSTP. Standard methods for collection and examination of wastewater were used to elucidate bacterial counts in the samples. The mean bacterial load at the influent was 7.1 x 107 CFU / 100 mL. Escherichia coli was the dominant bacterial type and the least common bacterial type was Vibrio parahaemolyticus. Other bacterial types found in the influent were Klebsiella aerogenes, Enterococcus faecalis, Salmonella typhi, Pseudomonas aeruginosa, Salmonella paratyphi, Vibrio cho/erae, Proteus mirabilis and Shigella flexneri. At the end of DSTP processing, the bacterial characteristic of the effluent was as follows: Enterococcus faecalis with mean total load of 2.7 x 104 ;:I: 1.7 x 103 CFU / 100 mL (Id = 0.0806), Escherichia coli estimated at 1.3 x 104 ± 1.1 x 103 CFU /100 mL iki= 0.1260), Pseudomonas aeruginosa measured as 2.5 x 102 ± 1.0 x 101 CFU /100 mL (Id = 0.1581), Salmonella typhi with load of 4.4 x 101 ± 5 x 10° CFU / 100 mL (Id = 0.1615) and Klebsiella aerogenes measured as 4.1 x 101 5 x 10° CFU / 100 mL (Id = 0.1897). In this regard, Enterococcus faecalis (Id = 0.0806) was the most resilient human pathogen. Seasonal variation in terms of quantity of bacteria was significant in both influent (F = 14.795, P = 0.001) and the effluent (F = 23.574, p = 0.000) with more bacteria found during the dry season. Bacterial load in the DSTP effluent showed diurnal variation with higher counts being found in the morning session (F = 22.788, p = 0.000). The bacteria types in Nairobi River, DSTP and Athi River were similar. The levels were higher in Nairobi River than in the DSTP effluent (F = 55.12, p = 0.001) and Athi River upstream (F = 13.638, p = 0.009). The performance of DSTP in terms of efficiency to remove bacteria from wastewater is below the set World Health Organization (WHO), Environmental Protection Agency (EPA) and National Environmental Management Authority (NEMA) guidelines for processed wastewater. The failure was replicated irrespective of the seasonal and diurnal variation. In conclusion, the DSTP and the Nairobi River portends a health risk to downstream communities and remedial intervention is urgently required.
Investigation of taint characteristics relating to the quality and flavour of coffee
(2012-06-07) Onani, Martin Opiyo; Ormand, K. L.
This thesis records the result of an attempt to substantiate an alleged complaint concerning the off-flavour in some batches of coffee as assessed by the trade liquorers of the Coffee Board of Kenya (CBK). The contribution potential of some of the volatile compounds derived from the fungicided used to control some pests and diseases was examined. These volatile substances are possibly produced during the biological breakdown of the fungicide and may be imparted to the coffee causing the taint. The methods of isolation of the volatile components of the fungicide and later the coffee samples were studied. The coffee samples were ground using a laboratory blender. The ground coffee samples were then extracted using 300ml of dichloromethane in a soxhlet apparatus for a period of eight hours. The excess solvent was evaporated in a rotary film evaporator and the concentrate stored at -20°C in the deep freezer. The conditions for the analysis of the extracts of the coffee samples were pre-determined. Analysis for the presence of the taint was done using the Sigma 3B gas chromatograph fitted with a fused silica capillary column, BPX5 (0.25), 25m X 0.32mm ID (SGE). The non-polar separatory phase was 5% phenyl equivalent modified siloxane. The carrier gas, hydrogen was used at a pressure of 6 pounds per square inch (PSI); the instrument was temperature programmed as follows: isothermal at 40°C for 1min, increased at a rate of 5°C/min to 190°C, and held for 5min. A Flame Ionization Detector (FID) was used. All the extracts were analysed using an on-column injection. Samples (0.025ml) were injected and chromatograms obtained. The retention times (Rt) from the chromatogram were compared to those of the standards run earlier and the results recorded. Analysis of the results supported the circumstantial evidence associated with the possible contribution of the fungicide prochloraz as responsible for the taint. The taint was observed in all the coffee samples from the areas where the fungicide was used. The fungicide Prochloraz is semi-systemic product and its degradation produces a number of metabolites including 2, 4, 6-trichlorophenol (TCP). Trichlorophenols are readily transformed to the corresponding trichloroanisole (TCA) in vivo. TCP has been shown in its own right to be a fungicide. This active ingredient could be the direct precursor of the TCA found in the tainted coffee samples. From this study there is a strong evidence that the fungicide Prochloraz produces the observed taint in the coffees studied. Continued usage of this fungicide should be dependant on the basis of further research, a check on application rates and taint appearance.
Mother's knowledge and practice of home care management of diarrhoea and risk factors in under 5 years in Nakuru, Kenya.
(2012-11-29) Mugo, Jackbeth M.; Orinda, G. O.; Anyango, S.; Otieno, Michael
Diarrhoea causes 1.8 million deaths each year worldwide and 90% of the deaths occur in children under 5 years. In less developed countries, a child has 6 to 8 episodes of diarrhoea per year. Kenya is one-amoung 15 countries in Africa and Asia that account for 75% of childhood diarrhoea deaths in children under 5 years. Kenya has an average of 4-5 episodes per child per year. At Nakuru general hospital, out of 1070 childhood diarrhoea admissions, there were 61 deaths in 2007. Policies such as Control of Diarrhoeal Diseases (CDD) 1993~ IMCI 1997, IMCIIHousehold Component 2000, Hand Washing Policy 2009 and Revised CDD 2010 were formulated to provide guidelines on the control and management of childhood diarrhoea yet diarrhoea disease continues to be a major cause of disease burden, Various programmes such as maternal child health (MCR), expanded programme on immunization (EPI), control of diarrhoea diseases (CDD) and integrated management of childhood illnesses (IMCI) were introduced in order to improve the quality of care and reduce morbidity and mortality and improve child survival, Studies have shown that mothers can give ORS at home, reduce hospital visits and therefore cut down on the treatment cost for diarrhoea disease. In Nakuru municipality in 2007, Baharini location diarrhoea was the second commonest cause of morbidity after pneuonia. Mothers were not able to provide appropriate treatment regimes leading to deterioration of the condition and delay in treatment. This is a barrier to achievement of MDG No.4 on child survival and achievement of Kenya's Vision 2030 of economic growth. This study was conducted in this peri-urban cosmopolitan area. The aim of the study was to assess the knowledge of mothers on signs of dehydration and complications of diarrhoea, determine the management regimes used by mothers during diarrhoea and identify household risk factors for diarrhoea transmission in children under 5 years of age in Baharini location, Nakuru municipality. Data was collected using pre-tested questionnaires and an observation checklist. Four hundred mothers of children under 5 years of age were interviewed. The respondents in the study were identified using simple random sampling where households/house blocks were selected using a table of digits. Data was analyzed using Chi-square test. The results showed that at the time of the study, there had been 41.5% diarrhoea episodes in the previous 6 months. Eleven point five percent (11.5%) of the respondents knew early signs of dehydration. Although 17.25% of the respondents knew that ORS was used to treat diarrhoea, only 7.75 % used ORS. Chi-Square test = 21.48 P value> 0.05 and the differences were considered significant. Eighteen point two percent (18.25%) of the respondents had adequate skill for preparing ORS. Carbohydrate foods were offered to children by 66% of the respondents but only 2% offered protein foods. It was observed that 51.75% of the households had poorly disposed domestic refuse although 73.25% of the respondents knew proper method of refuse disposal. Chi-square test = 34.44 and P value> 0.05, the differences were considered significant. Excreta was poorly disposed by 36.75% of the households. The study concluded that majority of the mothers had inadequate knowledge on signs of dehydration and complications of diarrhoea. Most mothers used inappropriate treatment regimes to manage diarrhoea. The methods used to dispose refuse and excreta in the household were high risk factors for diarrhoea transmission. The study recommended that the Clinicians should communicate appropriate information and skills to mothers on identification of early signs of dehydration and complications of diarrhoea and appropriate diarrhoea management regimes. The Ministry of Medical services should ensure that ORS sachets are available and accessible to mothers in the location. The Ministry of water should consider developing policies that promote chlorination of drinking water at the household level The department of public health should ensure that proper standards of sanitation in refuse and excreta disposal are maintained to reduce risk of diarrhoea transmission. The department should also consider methods of increasing excreta facilities in some of the estates such as Manyani and Lakeview.
Isolation and characterization of yeast from gallus gallus droppings in Kabigeriet village, Olenguruone
(2013-03-26) Kemoi, Edson Kipyegon
The Avian family has been a significant source of human epidemic. Over the last three decades, reports on yeast infections in humans have increased especially with respect to immunocompromised individuals. This is associated with increased morbidity and mortality especially in HIV/AIDS immunocompromised individuals. The purpose of this study was to isolate and characterize pathogenic yeasts from domestic Chicken (Gallus gallus) dropping. The droppings were collected from Kabigeriet Villages, Olenguruone Division, Kuresoi District and Nakuru County. The samples were collected from cages, houses and roosting sites. The samples (droppings and soil) were collected by swabbing or scooping fresh dropping from Chicken houses, grass, soil and trees using sterile plastic spoons, labeled and inserted in a zip lock safety bag. A total of Eighty four samples (dropping and soil enriched with chicken droppings) were sampled during the study. The samples were transported to the lab using a cool box. Processing of the samples was done at the Mycology laboratory, Centre for Microbiology Research, Kenya medical Research Institute. The samples were plated onto Typan blue agar and incubated at 37°C for two weeks checking was done daily. The isolates were identified according to morphological and biochemical characteristics. The droppings were tested for Cryptococcus by direct plating on Niger seed while Candida and Saccharomyces species by direct plating on Typan blue agar. Candida and Saccharomyces species were sub cultured on CHROM agar and Com meal agar for presumptive identification of various Candida species. Cryptococcus neoformans were sub cultured onto Christensen's urease agar. Geotrichum species were presumptively identified by lactophenol cotton blue. Analytical profile index test (API 20C AUX) was used for confmnation. Four types of yeasts were isolates; 35(57.4%) Candida species (9 Candida lusitanie, 7 Candida glabrata, 5 Candida albicans, 5 Candida tropicalis, 3 Candida parapsilosis, 2 Candida lipolytica and 2 Candida krusei) , 23(37.7%) Geotrichum candidum, 2(3.3%) Cryptococcus species (Cryptococcus neoformans -a-nd Cryptococcus lauren.ti) and 1(1.6%) Saccharomyces cerevisiae were isolated fromChickens dropping sampled. The results of this work demonstrated that domestic chicken (Gallus gallus) harbor Cryptococcus in their dropping and their close proximity to human habitation poses a risk of cryptococcal infection in HIV/AIDS immunocompromised persons. This could partly explain the high incidence of cryptococcosis in HIV/AIDS patients in Kenya.
Immune responses to cryptosporidium species in HIV/AIDS infected adults attending Kenyatta National Hospital, Nairobi, Kenya
(2013-07-30) Kanyi, Henry Muriuki
Cryptosporidium causes significant morbidity and mortality in AIDS patients worldwide particularly in developing countries where ART is not widely available or affordable. In Kenya, approximately 8% of the adult population lives with HIV/AIDS and Cryptosporidium has been reported as the leading indicator of death among adult HIV/AIDS patients. There is very little information about the molecular epidemiology of cryptosporidiosis and no documented data on immune responses to Cryptosporidium in HIV/AIDS patients in Kenya. A number of Cryptosporidium antigens have been found to induce host immune responses in immunocompetent persons, however, their effects in the immunity of immunocompromised individuals are not understood. Glycoprotein 15 (gp15) and Cryptosporidium 23 (Cp23) are conserved Cryptosporidium antigens that trigger host immune responses and are thus potent agents for vaccine development. This study was aimed at evaluating molecular epidemiology and immune responses against Cryptosporidium with an aim of preventing HIV/AIDS disease progression especially in immunocompromised HIV-Cryptosporidium co-infected persons. A total of 164 HIV/AIDS patients, 94p asymptomatic (no diarrhea) and 70 symptomatic (with diarrhea) respectively were recruited. Recruited persons provided stool samples for Cryptosporidium oocysts microscopic examination and blood for immune responses evaluation. Cryptosporidium species were identified through microscopy, and confirmed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) targeting 18S rRNA. Antibodies and IFN-γ responses against Cryptosporidium antigens were assessed by enzyme linked immunosorbent assays (ELISA). In this study, Cryptosporidium species was found to infect both symptomatic and asymptomatic persons. The species found were C. hominis, C. parvum, C. meleagridis, C. muris and C. Canis, with C. hominis being the most prevalent species. On immune responses there was a significant difference between patients with asymptomatic and symptomatic cryptosporidiosis in secretion of serum IgG to Chgp15 (P = 0.005) and Cp23 (P = 0.001) and fecal IgA to Chgp15 (P = 0.002). No significant difference was observed in IgM and IFN-g levels between patients with asymptomatic and symptomatic cryptosporidiosis. Cryptosporidium hominis was found to be associated with majority cases of Cryptosporidium infection in HIV/AIDS adult patients and often recurrent/chronic diarrhea in patients, especially those with low CD4 counts. Serum IgG to Chgp15 and Cp23 and fecal IgA to Chgp15 antibodies were also seen to be protective against diarrhea. These findings indicate that Serum IgG to Chgp15 and Cp23 and fecal IgA to Chgp15 antibodies may be good candidate antigens for vaccine development against Cryptosporidium species. However, study of the roles of CD8+ cells and other cytokines especially Th2 cytokines should be done to establish their effects on the host protective responses against Cryptosporidium observed in this study. These study findings indicate that improvement of immune status especially serum IgG and Innate mucosal IgA may be major remedial measures towards reducing effects of cryptosporidiosis which is a predominant infection in HIV/AIDS infected patients in Kenya.
Amoebiasis levels in relation to sanitation and personal hygiene among patients attending Kauwi and Muthale hospitals in Kitui county, Kenya
(2014) Mung’ang’a, Elijah Mativo
Entamoeba histolytica, an intestinal protozoan parasite, is the causative agent for amoebiasis which is the third leading parasitic disease causing deaths in humans after malaria and schistosomiasis. Globally, it is responsible for 40,000 to 100,000 deaths annually. Amoebiasis is on the rise in Kitui County and is responsible for diarrheal illnesses and early deaths in children under 5 years. There is no surveillance system in Kenya to combat the disease and its real level of occurrence remains unknown in Kitui west district. It occurs particularly in the tropics largely due to conducive temperatures for the parasite to complete its life cycle and is more common in areas with poor sanitation where barriers between human feces, food and water are inadequate. How this infection associated with age remains unclear. As a result, this study was aimed at determining the occurrence levels of E. histolytica infection among persons of various age groups attending Muthale mission and Kauwi sub-district hospitals in Kitui west district. The study involved observation of clinical signs and symptoms as well as microscopic examination of E. histolytica trophozoites and cysts in feces. The patients were sampled using simple random method. Fecal samples were collected from a total of 277 subjects attending the Kauwi sub-district and Muthale mission hospitals. To increase the chance of detecting the parasite in the stool, three direct smears were prepared from each sample and examined. Any observation of cyst or trophozoite was recorded. Formal-ether concentration technique was done for each sample that tested negative under direct smear and examined microscopically at low (20x) and high (40x) magnifications. The study collected data on personal hygiene and sanitation level in the study area by use of questionnaires. Laboratory data was obtained based on presence of cysts and / or trophozoites in the stool. Data on level of sanitation and infection by E. histolytica in different sexes and age groups were analyzed using Mean square contingency coefficient, Cramer’s V and Chi-square. The study revealed that 81 (29.2%) of the sampled individuals tested positive for amoebiasis of which 10.8% were males while females were nearly double at 18.4%. The most affected age group was the under five years at 58.8% and the least infected was 26 to 35 years at 20.6%. Source of water for domestic use played a key role in acquiring the parasite. Washing of hands, fruits and vegetables had no significance since the water used was from unsafe sources highly likely to have been contaminated with Entamoeba histolytica. This rendered fruitless the effort shown by the residents in maintaining good sanitation and personal hygiene. The current study findings are critical in preventing amoebiasis in the study area. This would have a long lasting impact in reducing the high child mortality rate in Kitui west district and other regions of the world with similar epidemiological factors. Thus, would put Kenya in an advantage position to achieve Millennium Development Goal number four and her Vision 2030.
The Role of Schistosoma mansoni Eggs in Immune Protection against Plasmodium berghei Infected Mice
(2014-02-17) Nyangahu, David Donald; Muturi, Margaret Wangui; Ochola, Lucy
The co-occurrence of malaria and schistosomiasis is common in tropical regions of the world. Malaria induces a strong Th1 response while schistosomiasis skews the response to a Th2. Several studies demonstrate a non consistent effect of schistosomiasis infection on progression of malaria. On one hand, schistosomiasis infections protect against cerebral malaria while on the other hand, they are associated with increased malaria severity. This study examined the role of Schistosoma mansoni eggs on Plasmodium berghei malaria progression in BALB/c mice. The objectives were to determine the changes in Th1, Th2 cytokines and IgG levels which are markers associated with malaria and schistosomiasis protection and also determine if S. mansoni eggs lead to protection from P. berghei malaria. Two groups of mice were used: the experimental group and the control group. The experimental group was injected with a triple dose of S. mansoni eggs at ten day interval before being challenged with P. berghei. The control group was infected with P. berghei only. Five mice from both groups were euthanized at each time point (day 3, 6, 9 and 12 post challenge with P. berghei) and the spleen and serum collected. Five mice from each group were monitored throughout the experiment. Parasitaemia was monitored daily using Giemsa stained blood smears. The results showed that the experimental mice exhibited lower levels of P. berghei parasitaemia (15.52%) as compared to the controls (23.06%). However the difference was not significant (p>0.05). IgG levels were found to be higher in the experimental mice compared to controls due to stimulation by soluble egg antigen (SEA). The differences in IgG levels between the two study groups was not significant (p>0.05). The levels of IFN- γ and IL-4 were higher in the experimental mice than the control group though the difference was not significant (p=0.213). The levels of IgG and IL-4 in experimental mice could be responsible for the delay in death reported in these mice and enhanced survivorship. In conclusion, S. mansoni eggs did not induce significant differences in cytokine and IgG levels; nevertheless they contributed to delaying death in the experimental mice by two days by enhancing levels of IgG and IL-4. These findings provide a pointer for further research in this field using higher animal model such as the non human primates for a better understanding of the immunomodulatory role of schistosoma eggs on progression of malaria.
Human Immunodeficiency Virus (HIV) Prevalence among Catha edulis forsk (Miraa) Users in Meru Region, Kenya
(2014-02-17) Njue, James Kinoti; Muchugi, Alice Njeri; Nyamache, A. K.
Khat or miraa (Catha edulis Forsk (Celastraceae) chewing is known to be a widespread habit in among selected communities in Kenya. Since HIV/AIDS was declared a national disaster in Kenya in 1999, the disease has become an obstacle to both health and development of the people. Nevertheless, the use of substance including C. edulis has dramatically increased despite the serious concern on control of HIV infection. This study was aimed at determining the HIV prevalence and impact of C. edulis chewing and social-demographic, knowledge, behavioral as risk factors to HIV infections and possible effects to CD4 and viral load counts among residents of Nyambene region of Meru County. A cross-sectional study was conducted among 267 individuals aged above15 years in the region during the period of May-December 2012. Data was collected using structured questionnaire and blood drawn from consenting participants. HIV status was determined by use of rapid tests; Determine and confirmed by ELISA test. CD4 and viral load counts were monitored (3months) for all HIV positive participants. The study established that the general HIV prevalence was 7.9% with women (8.1%) being affected more than men (7.6%) though not significant (p=0.019). Risk behaviours for HIV infection like C. edulis use were more observed among women engaged in Catha edulis business than men though not significant (p=1.468). Lack of knowledge on HIV transmission and prevention methods were associated with HIV status despite the high awareness of the disease. However there was significant difference on education level and breast feeding (p = 0.001), pregnancy (p=0.017) and HIV transmission during delivery (p=0.039). Most participants relied on radio as a source of information on HIV/AIDS which varied significantly with their education level (p = 0.001) and in HIV-TB co infection (p=0.005). There was significant difference on first CD4 count and the second, three months after (p = 0.001) unlike in viral load counts (p = 0.396). C. edulis use is risk behaviour for the spread of HIV infection. Ignorance, lack of knowledge and engagement into multiple sex partners predisposes people to risk of contracting HIV infection.
Care of prostate cancer patients in selected urology clinics in Nairobi County
(2014-07-28) Kinoti, K Fred Moses; Were, T.
Prostate cancer is an adenocarnoma of significant reproductive health burden in the world. Of the thirteen million cancer cases in the world, prostate cancer accounts for nine hundred thousand cases translating to 13.6% of the total cases and ranks as 5th most common cancer. The mortality rate among black population is 19 per 100,000 while among the Asian populations is 2.5 per 100,000.1n Africa, of the six hundred thousand cancer cases reported, prostate cancer accounts for thirty four thousand cases and is responsible for twenty four thousand deaths ranking 6th cause of death among men. In Kenya, twenty eight thousand cancer cases are reported and of these, prostate cancer accounts for 15.3% incidence of the total and causes a mortality of eight hundred and fifty Effective management of prostate cancer is largely dependent on early diagnosis, prompt treatment and follow up. However, due to lack of policy and adherence treatment protocols in Kenya, prostate cancer deaths have remained high. Therefore, the proposed descriptive cross sectional study will evaluate the care of prostate cancer patients in selected urology clinics within Nairobi County. The proposed study will embellish the existing discourse on prostate cancer management and practices and in addition, basic policy makers will have a reference point with regard to quality assurance and standard guidelines in prostate cancer treatment. A total of 156 patients and 18 urologists will be enrolled into the study. It will enroll all the patients with a diagnosis of prostate cancer and are undergoing treatment at the various urology clinics. Structured questionnaires will be used to collect social demographic data and data on availability and usage of prostate cancer protocols. Clinical information such as biometric values from the study participants will also be recorded in the questionnaires, Data will be cleaned and coded, dual entered in SPSS version 8.chi square will be used in determining associations between the urology characteristics and usage of protocols. Categorical data such as tumor grade and PSA levels will be summarized using contingency tables and bar charts, quantitative data will be summarized using proportions, medians, means, tabulated and graphed
Antinociceptive Activities of Acetone Leaves Extracts of Carissa Spinarum and Caesalpinia Volkensii in Mice
(Kenyatta University, 2016-04) Mworia, Joseph Kiambi
Despite the progress that has occurred in recent years in the development of therapy, there is still a need for effective and potent analgesics for pain. Pain is defined as unpleasant feeling essential for body‘s defense system. Pain is managed using analgesics such as aspirin,paracetamol, diclofenac, morphine, opioids, among others. Conventional antinociceptives are expensive and have many side effects. Continued use of these drugs may lead to tolerance. Medicinal plants have been used to relieve pain and form a better alternative. Herbal antinoceptives are affordable and have arguably fewer side effects. Carissa spinarum (Linn) is used to treat rheumatoid pain, fever and inflammation related disorders. Caesalpinia volkensii (Harms) has pharmacological activities that include antimicrobial,immune modulatory properties and antimalarial. These two plants are used locally by people in Embu County as analgesics.This study was designed to bioscreen the acetone leaves extracts of C. volkensii (Harms) and C. spinarum (Linn) for anti-nociceptive potential. The plant parts were collected from Siakago-Mbeere north sub-county, Embu County, Kenya. The samples were prepared and extraction of the active compounds carried out using organic solvent acetone in the ratio1:2.Swiss albino mice were divided into five groups of five mice each: Normal, negative, reference and experimental group.Pain was induced experimentally using formalin and acetic acid. The experimental groups were treated with 50 and 100mg/kg dose quantities of each plant extracts prepared. The acetone leaves extracts of the two plants were evaluated for antinociceptive properties in mice compared to the reference drug diclofenac sodium. Mice were injected intraperitoneally with doses of the herbs, diclofenac and the vehicle. Thirty minutes later the animals were injected with 0.01ml of 2.5 % formalin in the sub planter region of the left hind paw and the other set with 0.4ml of 5% acetic acid. The total time spent lifting; biting, licking the paw and writhing were counted and scored. The acetone leaves extracts lowered paw licking time in a dose dependant manner, The leaf extracts of C.volkensii at the dose levels of 50and 100mg/kg body weight reduced the formalin-induced pain in mice by 72.74% and 99.38 % respectively and acetic acid writhing by levels of 50 mg/kg body weight reduced the number of writhes by 81.40%, 100 mg/kg body weight did not reduce writhing. C.spinarum at the dose levels of 50mg/kg and 100mg/kg body weight reduced formalin induced pain by3.47%and 34.46 and 20.2% and 95.50% respectively. Acetic acid induced pain in mice by 73.77 % and 86.89 % respectively. Diclofenac reduced the pain by 15.34 in early phase and 98.02% in late phase. Further, the phytochemical screening results showed that the acetone leaves extracts of C.volkensii (Harms) and C.spinarum (Linn) have phytochemicals associated with anti-nociceptive activities. The study has established that the acetone leaves extracts of C.spinarum (Linn) and C.volkensii (Harms) are effective in management of pain. It is therefore recommended that further fractionation of the metabolites of the two plant extracts be carried out with a view to identifying the most active compounds for further development into drugs for management of pain and inflammation.
Prevalence of asymptomatic bacteriuria and antimicrobial susceptibility among women attending antenatal clinic of Gatundu Hospital in Kiambu County, Kenya
(Kenyatta University, 2018) Muturi, Ephantus Kiama
Asymptomatic bacteriuria in pregnancy is associated with risk of preterm birth and pyelonephritis if untreated. The apparent decline in immunity of pregnant women appears to promote the growth of both commensal and non-commensal microorganisms. Women with asymptomatic bacteria in the early pregnancy develop symptomatic bacteriuria later in pregnancy. The incidence of antibiotic resistance has been steadily increasing over the past few years resulting in limitation of therapeutic options. The aim of this study was to identify the prevalence of asymptomatic bacteriuria, the most common causative microorganisms and the antibacterial susceptibilities of the isolated microorganisms in pregnant women attending antenatal clinic at Gatundu Level Four hospital. An analytical cross sectional study design was applied to women with asymptomatic bacteriuria who attended the antenatal clinic and utilized the laboratory services. Quantitative data was analyzed using a computer package IBM SPSS statistics 20.0 by invoking the analysis of descriptive statistics and focusing on frequencies and percentages. Pearson Chi square test was carried out where p< 0.05 values were considered significant. A total number of 120 asymptomatic pregnant women were screened for ASB by urine culture using a semi quantitative culture method. Midstream urine was collected and aerobically incubated at 37° C on CLED and MacConkey Agar. Growth of >1x105 CFU/mL was taken as significant bacteriuria. Gram-negative bacteria were identified and confirmed by biochemical tests. In this study, the prevalence of bacteriuria among the pregnant women was 10.8 %. The prevalences of isolated bacteria from urines of pregnant women were: Escherichia coli 69.2 %, Proteus species 23.1 % and Klebsiella species 7.7 %. Among cases which showed positive cultures, 10.0 % were nulliparous and 11.4 % were multiparous. The highest incidence (33.3 %) of asymptomatic bacteriuria was found in the maternal age category age group of 31-35 years. Majority of the isolates in the study were susceptible to Cefataxime (100 %) Ofloxacin (88.9 %) and Gentamycin (44.4 %). Findings of the study provide empirical data on the prevalence of asymptomatic bacteriuria within pregnant women. The prevalence of asymptomatic bacteriuria is high among this study population, therefore all pregnant women should be screened during their first visit to ante-natal clinic by urine culture to detect and treat ASB to avoid complications that may ensue in both mother and fetus. Establishment of antibiotic policy to guide against the emergence of resistant organisms that cause asymptomatic bacteriuria should be effected.
Shigella Serogroups Distribution, Antimicrobial Susceptibility and Risk Factors of Shigellosis Among Children Presenting with Diarrhoea at Banadir Hospital in Mogadishu, Somalia
(Kenyatta University, 2022) Ali, Bilan Sheikh; Nelson Menza
Bacillary dysentery (shigellosis) is a public health concern globally, ranking second among the top killer diseases in children aged five years and below. The burden of shigellosis is disproportionately higher in middle and low-income countries due to poor sanitation and insufficient clean drinking. With shigellosis treatment facing unprecedented challenges arising from multidrug-resistant (MDR) Shigella strains, understanding the local epidemiology is vital to inform prevention and control interventions. This study determined Shigella serogroups distribution, antimicrobial susceptibility (AS) patterns and shigellosis associated factors among children with diarrhoea at Banadir Hospital, Somalia. The study design was cross-sectional, and participants purposively recruited to include children aged five years and below with diarrhoea, whose parent/guardian gave formal consent. With participants sociodemographic and clinical data sought using a questionnaire, 180 stool and rectal swab samples were collected and processed following the standard microbiological methods. Shigella serogroups AS patterns were elucidate using the Kirby- Bauer method. The data were generated and presented in tables, and the association between the shigellosis, socio-demographic and environmental characteristics of the participants, was computed using the Chi-square (χ 2 ) and Pearson correlation statistics using IBM SPSS version 23 software for Windows. The children majority were females 111/180 (61.7%), while males accounted for 38.35% (69/180) of the total samples collected. The children mean age was 20.1 months with 12.87 months standard deviation (SD). Generally, shigellosis prevalence among children with diarrhoea was 20.6% (37/180). While S. boydii was absent among the bacteriologic cause of bacillary dysentery, the other three (3) Shigella serogroups were isolated; S. flexneri, S. sonnei and S. dysenteriae type 1 (shiga bacillus). Of these, the predominant serogroup was Shigella flexneri (26/37, 70.3%) and the least prevalent was shiga bacillus (5/37, 13.5%) (t = 15.187; p = 0.0001). There was 100% resistance to ampicillin, trimethoprim-sulfamethoxazole, and tetracycline among the Shigella serogroups isolated. Except for S. dysenteriae type 1, which remained susceptible to ciprofloxacin, 19.2% (5/26) of S. flexneri and 50% (3/6) of S. sonnei isolates showed resistance. Forty per cent of the Shiga bacillus were ceftriaxone (CRO) nonsusceptible. Shigella sonnei had the highest CRO resistance (66.7%, 4/6) and the least resistant was S. flexneri (38.5%, 10/26). All the serogroups were susceptible to azithromycin. MDR phenotype AMP/SXT/TE (37/37, 100%) was the most dominant, and the least prevalent was AMP/SXT/TE/CIP/CRO (8.1%, 3/37). There was no significant association of shigellosis among children aged five years and below with the risk factors investigated in Banadir Hospital. This study showed high shigellosis prevalence, and S. flexneri and Shiga bacillus as the most and least prevalent serogroup, respectively, among children (≤ 5 years) in Banadir Hospital. CIP and CRO resistant Shigella, including MDR strains, have emerged among children (≤ 5 years) in Mogadishu, Somalia. Therefore, prevention and control interventions are urgently required to mitigate MDR Shigella associated infections in Banadir Hospital and the surrounding regions.
Optical Characterization of Dye-Sensitised Tio2 Thin Films for Solar Cell Applications
(Kenyatta University, 2022) Kainyu, Kabuga Pauline; W. K. Njoroge
The use of dye-sensitized solar cells to harness solar energy has gained interest because of their simplicity and are cheaper to manufacture. They can also be glazed on flexible substrates for improved conversion efficiency. Standard DSSC consist of materials made of transparent conducting oxides (TCOs) with titanium dioxide (TiO2) used as semiconducting material. Synthetic dyes or natural dyes are utilized as sensitizers to modify the TiO2 film. The goal of the current study was to find out the potential of organic dyes from blue berry and spinach as sensitizing agents for TiO2 films by investigating their influence on optical properties of the films and energy conversion efficiency of DSSC employing the said films. The blueberry and the spinach dyes were extracted from blueberry fruit and spinach leaves respectively and were taken into the nano-porous titania substrates. The nano-porous TiO2 paste was sourced from Solaronics SA and its film generated on the glass surface by employing screen printing technique. Ruthenium dye was used as a reference dye. Using DUV3700 spectrophotometer the TiO2 film was characterized to obtain its optical properties which were transmittance, reflectance and absorbance for both sensitized and un-sensitized TiO2 films. The optical results showed blue berry sensitized TiO2 had low reflectance, transmittance and high absorbance compared to all other samples both sensitized and un-sensitized. Spinach sensitized TiO2 films had high reflectance, transmittance and low absorbance. On energy conversion efficiency, the DSSC fabricated using Ruthenium dye had the highest efficiency of ~ 2%. The DSSC fabricated with Blue berry dye followed closely at ~ 1.5 % efficiency while the DSSC employing spinach dye had the lowest conversion efficiency of ~ 0.6 %. Diluting the dyes with distilled water improved conversion efficiency for DSSCs fabricated employing the blue berry and spinach dyes. From the study blue berry dye has shown a more promising potential for fabrication of the DSSCs. However, further study is required to optimize the concentration of the dyes as diluting the dyes enhanced the performance of the dye as a DSSC sensitizer.
Teratogenic and Toxicity Effects of Dichloromethane-Methanol Extracts of Toddalia Asiatica (L) Lam and Acacia Polyacantha Plants from Subukia, Nakuru County, Kenya
(Kenyatta University, 2023-04) Gichuru, Joseph Mwangi; John K. Mwonjoria; Nelson C. Menza; David N. Mburu
Abstract
Method Verification and Quality Assurance of Quantitative Analytical Methods in Clinical Chemistry Department, Machakos Level V Hospital, Machakos County, Kenya.
(Kenyatta University, 2024-04) Mutie, Charles Kyalo
Kenya seeks greater accreditation of medical labs for accurate results and mutual recognition. Quality management systems and competence are essential in public health laboratories. Machakos Level V hospital laboratory is accredited to ISO 15189:2012. Accreditation leads to accurate and mutual recognition of test results. The facility has state of the art equipment found in other leading hospital laboratories worldwide and several publications on performances. Accreditation requires method validation and verification studies. No guidelines exist for verifying methods at set intervals, which is crucial for providing quality healthcare. Verification and quality assurance of analytical methods follow standard procedures to improve accuracy, patient safety, and lab efficiency. This study aimed to improve the frequency of method verification in the Clinical Chemistry department at Machakos level V Hospital by analysing accuracy, precision, and quality assurance. Test methods were assessed for accuracy, precision, and quality assurance using quality control material. Precision and accuracy were measured and compared to the manufacturer's ranges. Samples were processed on time and External Quality Assurance results were analysed for enhancement opportunities. External Quality Assurance score goal was 80%. Failure to achieve it led to a nonconformance event that necessitated root cause analysis and corrective action. Analysis was conducted on 13 chemistry analytes at Machakos Level V Hospital Laboratory in Kenya. Results were based on internal quality control data. On short term precision twenty-five out of twenty-six analytes met the Clinical Laboratory Improvement Amendments target coefficient of variation %, except for chloride normal control. Fifteen out of twenty-six analytes met manufacturers recommended coefficient of variation % while eleven failed. Parameters that failed include aspartate aminotransaminase, direct bilirubin, total bilirubin, chloride, potassium, sodium, total protein, and urea. Long term precision results revealed twenty analytes level met target Coefficient of Variation %, but three failed both controls that is chloride, potassium, and urea. None of the thirteen analytes met manufacturers recommended coefficient of variation % hence failure. All the analytes passed accuracy with a mean within manufacturer's verification interval and Z-score of less than ±1.96. Results were consistent with manufacturer recommendations for all analytes indicating no statistical difference. Total error was calculated based on Clinical Laboratory Improvement Amendments guidelines. All analytes passed except chloride normal control. On quality assurance, a retrospective analysis on thirteen general chemistry analytes spanning August 2021 to February 2023 was conducted. Review of internal quality control found violations of Westgard rules, inconsistent and insufficient reviews by the laboratory, they reviewed three out of twelve months. Data on performance from European Society for External Quality assessment was utilized. Passes were achieved in November 2022, August 2022, and March 2022 cycles, indicating consistent improvement. Failure was recorded in May 2022, November 2021, and August 2021. Results were shared with management for corrective action and preventive action. The findings will aid other counties, accreditation bodies and the Ministry of Health in refining policies crucial for laboratory accreditation.

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