应用复合PCR方法检测沙门菌的研究

参照文献报道的沙门菌Repeatse和hisJ基因片段的引物序列 ,设计并合成了 2对引物 ,对沙门菌属和非沙门菌属的标准菌株及分离菌株进行了复合PCR扩增反应。结果 ,沙门菌PCR产物均出现 199bp与 4 95bp的特异性DNA扩增条带 ,而非沙门菌均未出现扩增条带 ,证明这 2对引物具有沙门菌属特异性。将提取的沙门菌DNA做梯度稀释 ,测定该复合PCR体系的敏感度。结果表明 ,此体系可检出 10 3CFU/mL菌液提取的DNA模板。应用上述方法 ,对进境鱼粉阳性样品进行了检测 ,均出现阳性结果。本研究表明 ,复合PCR是一种特异、敏感、快速的沙门菌检测方法 ,可应用于口岸系统鱼粉、肉骨粉的日常检测。

Study on detection of Salmonella by multiplex PCR

A multiplex PCR method was established for detection of Salmonella by using two pairs of primers unique to Repeat sequence and hisJ gene segments of Salmonella species. Two specific DNA amplicons were present in the standard strains and laboratory isolates of Salmonella, but absent in the non-(Salmonella) samples, which showed that two pairs of primers are Salmonella-specific. The sensitivity of this method was confirmed by serial dilutions of extracted SalmonellaDNAs while approximately (10~(3))colony forming units per 1 mL (CFU/mL) were detected. Moreover, positive samples, indicated by conventionalmicrobiological assays followed by confirmative serological tests in about seven days, were also demonstrated by the multiplex PCR within hours. It is revealed in this study that the multiplex PCR, specific, sensitive, rapid, can be applied to routine test of imported fish meal, meat and bone meal.