Comparison of short-term and long-term protocols for stabilization and preservation of RNA and DNA of Leishmania, Trypanosoma, and Plasmodium

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dc.contributor.authorNgeranwa, J.J.N.
dc.contributor.authorBasiye, Frank L
dc.contributor.authorSchoone, Gerard J
dc.contributor.authorMinnaar, Marcel Beld, Rene
dc.contributor.authorWasunna, Monique K
dc.contributor.authorSchallig, Henk D. F. H.
dc.date.accessioned2013-05-07T08:57:31Z
dc.date.available2013-05-07T08:57:31Z
dc.date.issued2010-08-18
dc.description10.1016/j.diagmicrobio.2010.08.018 pp.66-73en_US
dc.description.abstractMolecular tools continue to be important in the prevention and control of parasitic diseases. However, using these techniques directly in the field remains a major challenge. Therefore, the preservation of clinical samples collected from endemic field areas for later analysis remains an important preanalytical process. This study aimed at identifying a suitable protocol for stabilization and preservation of RNA and DNA in bioclinical specimens for Trypanosoma, Leishmania, and Plasmodium research. Both spiked and unspiked blood samples were preserved in 7 protocols (different media; storage temperatures). Samples were evaluated for possible degradation of DNA and RNA along the storage duration up to the 10th week. Nucleic acid targets were assessed as follows: (i) Trypanosoma and Plasmodium RNA analysis was done using real-time nucleic acid sequence-based amplification (RT-NASBA) for 18S rRNA and for stage-specific Pfs25 mRNA, respectively; (ii) Trypanosoma DNA assessment analysis was conducted by using a conventional PCR for 18S rDNA; (iii) Leishmania RNA analysis was performed with a quantitative NASBA for 18S rRNA and Leishmania DNA assessment with an RT-PCR for 18S rDNA. Findings suggested that a newly developed L3™ buffer proved to be reliable and suitable for both short- and long-term preservation of parasite nucleic acid material. This buffer is envisaged to be suitable for utilization in field situations where resources are limited.en_US
dc.identifier.citationDiogonistic Microbiology and Infectious Disease (impact factor: 2.45). 01/2011; 69(1):66-73.en_US
dc.identifier.urihttp://ir-library.ku.ac.ke/handle/123456789/6761
dc.language.isoenen_US
dc.publisherDiogonistic Microbiology and Infectious Diseaseen_US
dc.titleComparison of short-term and long-term protocols for stabilization and preservation of RNA and DNA of Leishmania, Trypanosoma, and Plasmodiumen_US
dc.typeArticleen_US
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