动物性食品源弓形菌23S rRNA基因PCR检测方法的建立

Establishment of PCR Method for Detecting Animal Food-Borne Arcobacter Based on the 23S rRNA Gene

建立了一种快速、准确检测弓形菌(Arcobacter)的PCR方法。根据弓形菌23S rRNA基因序列设计引物,对弓形菌标准菌株、弓形菌食品分离株及非弓形菌属菌株进行PCR扩增。结果表明,弓形菌标准菌株和35株弓形菌食品分离株扩增后均可得到688 bp的目的条带,11株非弓形菌属菌株均未见任何扩增条带,对弓形菌的最低检测限为1.12×103CFU/mL。该方法操作简单、检测周期短、灵敏度高、特异性好,可用于食品中弓形菌的快速检测。

A rapid and accurate method of polymerase chain reaction （PCR） for detecting Arcobacter species was established.A pair of primers was designed by using the 23S rRNA gene as objective sequences to amplify the genes of reference Arcobacter,isolated Arcobacter and non-Arcobacter strains.Expected fragments of 688 bp were obtained for reference Arcobacter and 35 Arcobacter food isolates.No specific fragment was detected in the 11 strains of the non-Arcobacter bacteria.The result of specificity tests showed that the detection limit could reach 1.12×103 CFU/mL.The PCR procedure is a sensitive and specific method and can be used to rapidly detect the pathogenic Arcobacter in food.