Direct Detection of Shigella flexneri and Salmonella typhimurium in HumanFeces by Real-Time PCR
Direct Detection of Shigella flexneri and Salmonella typhimurium in HumanFeces by Real-Time PCR
- 주제(키워드) Shigella flexneri , Salmonella typhimurium , real-time PCR , human fecesShigella and Salmonella infections are generally diagnosedfecal cultures. Culture methods are precise , but relativelyinefficient , time-consuming , and labor intensive. Polymerasecha
- 발행기관 한국미생물·생명공학회
- 발행년도 2007
- 총서유형 Journal
- UCI G704-000169.2007.17.10.009
- KCI ID ART001090035
- 본문언어 영어
초록/요약
We have established a SYBR Green-based real-time PCR method using AnyDirect solution, which enhancesPCR from whole blood, for direct amplification of the virAgene of Shigella flexneri and the invA gene of Salmonellafrom human feces without prior DNA purification.When we compared the efficiency of conventional or real-time PCR amplification of the virA and invA genes from thesupernatant of boiled feces supplemented with S. flexneri andS. typhimurium in the presence or absence of AnyDirectsolution, amplification products were detected only in reactionsto which AnyDirect solution had ben added. The detectionlimit of real-time PCR was 1×104 CFU/g feces for S. flexneriand 2×104 CFU/g feces for S. typhimurium; this sensitivitylevel was comparable to other studies. Our real-time PCRasay with AnyDirect solution is simple, rapid, sensitive, andspecific, and allows simultaneous detection of S. flexneri andS. typhimurium directly from fecal samples without priorDNA purification.
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