Carbapenemase-producing Enterobacteriaceae(CPE) is a global health issue due to their hasty dissemination through the transfer of carbapenemase genes. Hence, rapid detection is necessary to take relevant control measures against CPE infections/colonization. We established a rapid and multiplex CPE detection system - Single Tag Hybridization Printed Array Strip (STH-PAS) by targeting the four different major carbapenemases. STH-PAS is a DNA-DNA hybridization technique where the oligonucleotide tag in the primer of PCR product hybridizes to its probe imprinted on a chromatographic strip without denaturation. Further, the efficacy of STH-PAS in detecting CPE directly in clinical samples is evaluated.
STH-PAS was tailored to detect various alleles of the four carbapenemase genes – NDM, KPC, IMP, and OXA-48 like in a single reaction. Then, the efficiency of hybridization in STH-PAS for detection of carbapenemases was compared with conventional PCR. The efficiency of carbapenemase detection by STH-PAS was analysed in CPE (n=49) and non-CPE strains (n=10). A total of 114 CPE suspected stool samples were subjected to STH-PAS to examine its utilization for direct clinical samples.
The ideal conditions for hybridization without non-specificity in STH-PAS was determined. STH-PAS was found to be 10 times more sensitive than conventional PCR techniques. It showed both sensitivity and specificity of 100% for carbapenemase detection in bacterial strains (n=59). As it is not affected by any of the inhibitory substances in clinical specimens, STH-PAS showed 90.91% sensitivity and 98.57% specificity in detecting carbapenemase directly in stool samples (n=114).
The results of the current study show that STH-PAS possesses several advantages as a good detection system for CPE. As it is very rapid and simple to interpret the results with naked eye, STH-PAS could be applied in poorly resourced countries. It has been planned to assess the effectiveness of STH-PAS as a surveillance tool in clinical settings to control the transmission of CPE.
R. K. Shanmuga Kani,
N. Yamamoto, None
N. Sakamoto, None
H. Hagiya, None
H. Yoshida, None
T. Kodera, None
P. Santanirand, None
S. Hamada, None
K. Tomono, None
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