| dc.description.abstract | Resistance to β-lactam antibiotics through β-lactamase production by Enterobacteriaceae continues to burden the health-care
sector worldwide. Traditional methods for detection of β-lactamases are time-consuming and labor-intensive and newer methods
with varying capabilities continue to be developed. The objective of this study was to develop a multiplex PCR (M-PCR) system
for the detection of blaSHV, blaTEM, blaCTX-M-1, blaCTX-M-9 and blaOXA-1 group genes and to apply it in clinical Klebsiella
pneumoniae and Escherichia coli strains. To do this, we used group-specific PCR primers in singleplex reactions followed by
optimization into multiplex reactions. Specificity and sensitivity of the M-PCR were then evaluated using 58 reference strains
before its application to detect bla group genes in 203 clinical Enterobacteriaceae strains. PCR amplicons were sequenced to
determine the β-lactamase subtypes. The M-PCR system exhibited 100% specificity and sensitivity. In all, 83.7% of
K. pneumoniae and 89.8% of E. coli clinical strains harbored bla group genes with 46.9%, 40.1%, 15.0%, 21.1% and 6.1%
of K. pneumoniae having blaSHV, blaTEM, blaCTX-M-1, blaCTX-M-9 and blaOXA-1 group genes, respectively, whereas 12.2%, 77.6%,
22.4%, 36.7% and 8.2% of E. coli had blaSHV, blaTEM, blaCTX-M-1, blaCTX-M-9 and blaOXA-1 group genes, respectively. BlaSHV-1,
blaSHV-11, blaSHV-27, blaSHV-33, blaSHV-144, blaTEM-1, blaTEM-135, blaOXA-1, blaCTX-M-3, blaCTX-M-9, blaCTX-M-14, blaCTX-M-15,
blaCTX-M-27, blaCTX-M-55, blaCTX-M-65 and blaCTX-M-104 were detected. In conclusion, the M-PCR system was efficient and versatile
with an advantage of simultaneously detecting all the targeted bla group genes. Hence, it is a potential candidate for developing
M-PCR kits for the screening of these genes for clinical or epidemiological purposes. | en_US |
| dc.description.sponsorship | This work was supported by grants from National Natural Science Foundation
of China (NSFC) (81101300, 31370164, J1103609 and J1210062), Heilongjiang
Educational Agency (1155G34), Heilongjiang Provincial Science and
Technology Innovation Team in Higher Education Institutes for Infection and
Immunity, Harbin Medical University | en_US |
