豬沙門氏菌的PCR 與RFLP-PFGE 識別技術比較研究   全文下載

Evaluation of PCR-based Fingerprinting Comparatively to the RFLP-PFGE for Discrimination of Salmonella sp. Isolated from Slaughtered Pork

有效地進行沙門氏菌流行病的監控需要應用快速和準確的基因測序方法技術。為了評價PCR 和RFLP-PFGE 測序方法的差異性，本文基於快速的PRC 方法（ERIC-PRC、M13-PRC 和RAPD 方法）研究比較了73 個沙門氏菌種。結果表明：所用引物的效果是不相同的，不同血清類的引物其性能指標也不同。在血清水準以外，具有OPC19 的ERIC-PCR 和RAPD 的引物對沙門氏菌的識別是無效的。相反地，M13-PCR、OPC15-RAPD 和OPB17-RAPD 引物在血清內能識別沙門氏菌，但比RFLP-PFGE 的效果要差，表明RFLP-PFGE 不是沙門氏菌流行病監測的唯一測序方法。在沙門氏菌種的篩選過程中，PCR 方法比RFLP-PFGE 方法更快和經濟，且更具有吸引力的一種方法。

Effective epidemiological surveillance and control of Salmonella sp. requires accurate and expeditious genetic typing methods. In the present study, rapid PCR-based methods (ERIC-PCR, M13-PCR and RAPDs) were applied to 73 Salmonella sp. isolates, and the results compared with those previously obtained by RFLP-PFGE (Salmonella gold standard genotyping method), in order to evaluate their discriminatory ability. Results were very diverse among the primers used and, for each primer, the performance level was variable among the different serotypes. ERIC-PCR and RAPD with OPC19 was inefficient for Salmonella sp. discrimination beyond the serotype level. In opposite, M13-PCR, OPC15-RAPD and OPB17-RAPD allowed intraserotype discrimination that, in general, were less discriminative than RFLP-PFGE, indicating that should not be used as a unique typing method in epidemiological studies. Nevertheless, in particular situations, these PCR methods, which are faster and less expensive than RFLP-PFGE, could offers an attractive choice as a preliminary screening of the isolates to reduce the number of suspicious isolates that should be subsequently typed with a more discriminative and accurate methods such as RFLP-PFGE.