장티푸스균 신속진단을 위한 multiplex PCR 진단법 확립
Further Evaluation of Multiplex PCR for Rapid Detection of Salmonella Typhi
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Typhoid fever is a class I legally designated communicable disease in Korea; and if remains as an important public health problem in many developing countries. It takes at least 3-5 days to detect and identify Salmonella Typhi (S. Typhi) by classical diagnostic method. For this reason, multiplex PCR (mPCR) was evaluated in detecting and identifying S . Typhi. In this study, forty-three bacterial strains, which consisted of 42 Salmonella enterica serovars and one Citrobacter freundii . were used to evaluate the promptness of mPCR in detecting and identifying S . Typhi. mPCR was performed with four genes which were known for representing Salmonella spp and/or S . Typhi: invA, fliC-d, viaB and prt. invA and prt gene was amplified in all strains and viaB gene was in only S. Typhi. fliC-d gene was amplified in three serovars: S . Typhi, S. Schwarzengrund and S. Livingstone. After specificity test,mPCR was modified as triplex PCR with three genes (invA, fliC-d, and viaB) and the sensitivity test was performed against S. Typhi-inoculated stool samples. mPCR was able to detect S. Typhi cell suspension of 1×105 cfu/mL. We found that modified multiplex PCR was useful to detect S. Typhi from stool samples within 24h whereas it takes 3-5days to detect by classic diagnosis method.
Abstract
Typhoid fever is a class I legally designated communicable disease in Korea; and if remains as an important public health problem in many developing countries. It takes at least 3-5 days to detect and identify Salmonella Typhi (S. Typhi) by classical diagnostic method. For this reason, multiplex PCR (mPCR) was evaluated in detecting and identifying S . Typhi. In this study, forty-three bacterial strains, which consisted of 42 Salmonella enterica serovars and one Citrobacter freundii . were used to evaluate the promptness of mPCR in detecting and identifying S . Typhi. mPCR was performed with four genes which were known for representing Salmonella spp and/or S . Typhi: invA, fliC-d, viaB and prt. invA and prt gene was amplified in all strains and viaB gene was in only S. Typhi. fliC-d gene was amplified in three serovars: S . Typhi, S. Schwarzengrund and S. Livingstone. After specificity test,mPCR was modified as triplex PCR with three genes (invA, fliC-d, and viaB) and the sensitivity test was performed against S. Typhi-inoculated stool samples. mPCR was able to detect S. Typhi cell suspension of 1×105 cfu/mL. We found that modified multiplex PCR was useful to detect S. Typhi from stool samples within 24h whereas it takes 3-5days to detect by classic diagnosis method.
참고문헌 (11)
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