活的非可培养状态沙门菌RT-PCR检测
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国家质检总局科技计划项目(2009IK168);“十二五”吉林省教育厅科学技术研究项目(201148)


RT-PCR for Viable but Non-culturable Salmonella
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    摘要:

    依据沙门菌 Salmonella 活的非可培养状态(VBNC)差异基因序列设计特异性引物C1和C5,并对处于VBNC的沙门菌模型开展RT-PCR检测.结果表明,引物C1和C5可从模型中分别扩增出198和137 bp的目的片段,这些片段与差异基因的核苷酸序列相似性均大于99%.而且所获得的电泳图谱也不同于正常状态的沙门菌、霍乱弧菌 Vibrio cholerae 等一些食源性致病菌.另外,2对引物的最低cDNA检出量分别为30.65和3.065 pg/μL.由此证明,引物C1和C5具有较好的特异性和敏感性,可用于建立VBNC沙门菌的检测方法.

    Abstract:

    Viable but non-culturable Salmonella was detected by RT-PCR with primers designed according to the differential viable but non-culturable (VBNC) genes, ybbB and rh1B. The results showed that primers C1 and C5 respectively amplified fragments about 198 and 137 bp, their nucleotide sequence homology with ybbB and rh1B was both over 99%, and the electrophoretogram form VBNC Salmonella was different from that of normal Salmonella, Vibrio cholerae, et al. The lowest cDNA detection level with primers C1 and C5 in VBNC Salmonella was respectively 30.65 and 3.065 pg/μL. Primers C1 and C5 with specificity and sensibility were applied to establish the detection method in VBNC Salmonella.

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李 影,王伟利,孟庆峰,钱爱东.活的非可培养状态沙门菌RT-PCR检测[J].华南农业大学学报,2013,34(1):98-100

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  • 收稿日期:2012-07-20
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  • 在线发布日期: 2013-02-09