CAJ | 학술논문

根据弯曲菌16S rRNA基因的靶序列设计特异性引物和探针，对反应条件和试剂浓度进行优化，建立了快速检测鸡肉中弯曲菌的荧光定量PCR方法，并对其特异性、敏感性和重复性进行评价。该方法除了对弯曲菌有扩增曲线外，对其他常见食源性病原菌均未有扩增曲线，具有较好的特异性；弯曲菌的最低检测限为10 CFU/mL；重复性实验获得标准曲线相关系数为R2=0.999，批内可重复性变异系数在0.12%-2.1%之间，批间可重复性变异系数为1.7%。应用该方法对68份鸡肉样品检测弯曲菌阳性率为95.6%（65/68），传统分离方法阳性率为89.5%（60/68），两种方法的阳性符合率为89.7%。本研究建立的荧光定量PCR方法灵敏度高、特异性好、操作简单，大大缩短检测周期，可作为检测鸡肉样品中弯曲菌的手段，为鸡肉样品弯曲菌的流行病学调查研究提供新的工具。
근거만곡균16S rRNA기인적파서렬설계특이성인물화탐침，대반응조건화시제농도진행우화，건립료쾌속검측계육중만곡균적형광정량PCR방법，병대기특이성、민감성화중복성진행평개。해방법제료대만곡균유확증곡선외，대기타상견식원성병원균균미유확증곡선，구유교호적특이성；만곡균적최저검측한위10 CFU/mL；중복성실험획득표준곡선상관계수위R2=0.999，비내가중복성변이계수재0.12%-2.1%지간，비간가중복성변이계수위1.7%。응용해방법대68빈계육양품검측만곡균양성솔위95.6%（65/68），전통분리방법양성솔위89.5%（60/68），량충방법적양성부합솔위89.7%。본연구건립적형광정량PCR방법령민도고、특이성호、조작간단，대대축단검측주기，가작위검측계육양품중만곡균적수단，위계육양품만곡균적류행병학조사연구제공신적공구。
To establish a rapid assay for quantitative detection of Campylobacter spp.,a pair of primers and fluorescent probe were designed according to 16S rRNA sequence of Campylobacter spp.,and then a real-time PCR method was successfully established to detect Campylobacter spp. Results demonstrated that the method was speciifc for ampliifcation of Campylobacter spp.,but no ampliifcation for other related bacteria. The detection limit of the assay was 10CFU/mL and the correlation coefifcient of standard curve(R2)was 0.999. The detection of 68 chicken meat samples showed that the positive rate was 95.6%,while the positive rate was 89.5% by traditional culture method. The method was simple,rapid,speciifc and sensitive for rapid detection of Campylobacter spp. and a new tool for epidemiological investigation of Campylobacter spp. in chicken meats.