焦磷酸测序检测食品中鼠伤寒沙门氏菌

摘要

According to the specific sequence of Salmonella typhimurium, amplification primers and sequencing primers were respectively designed to establish the pyrosequencing method in this study. Target fragments were ampli-fied with PCR by the specific primers aiming at S. typhimurium. And prepare single-stranded templates and carried out pyrosequencing with sequencing primers. The sequencing results showed that 6 strains of S. typhimurium from dif-ferent sources could all be amplified as TACAACCGGA GTGCACATTA ATCCCGCAGC gene fragments, while 30 strains of the negative control strains could not be amplified. BLAST comparison indicated that the sequences were 100% match with the base sequence of S. typhimurium in GenBank. The pyrosequencing method is a rapid, accurate detection method, it could be used for rapid detection of S. typhimurium in food.%根据鼠伤寒沙门氏菌的特异序列,分别设计扩增引物和测序引物,建立焦磷酸测序检测鼠伤寒沙门氏菌的方法。针对鼠伤寒沙门氏菌设计特异性扩增引物,对目标片段进行PCR扩增,然后制备单链模板,并利用测序引物进行焦磷酸测序。测序结果表明,6株不同来源的鼠伤寒沙门氏菌均可以扩增出碱基序列为TACAACCGGA GTGCACATTA ATCCCGCAGC的基因片段,而30株阴性对照菌株均未得到扩增。进行BLAST比对表明,该序列与GenBank中鼠伤寒沙门氏菌的碱基序列100%匹配。焦磷酸测序法是一种快速、准确的检测方法,可用于食品中鼠伤寒沙门氏菌的快速检测。