摘要
目的:建立快速检测沙门氏菌、无乳链球菌和金黄色葡萄球菌的多重PCR方法。方法:根据沙门氏菌(Salmonella)组氨酸转运操纵子基因、无乳链球菌(Streptococcus agalactiae)的STRA-Agl-23-1D基因和金黄色葡萄球菌(Staphylococcus aureus)的耐热核酸酶nuc基因分别设计引物,进行PCR扩增及反应条件的优化,建立这3种菌的多重PCR检测方法。结果:通过建立多重PCR方法,3对引物同时特异性地扩增出495 bp、360 bp和279 bp目的片段;3种菌的检测限:沙门氏菌1.9×102 cfu/mL、无乳链球菌2.0×102 cfu/mL、金黄色葡萄球菌2.9×102 cfu/mL;3种菌DNA含量的(最低)检出限分别为3.86、25.5、3.47pg。结论:本文建立的多重PCR检测方法,简单、快速、灵敏度高,具有很好的应用前景。
Objective：To establish a multiplex polymerase chain reaction（M-PCR） method for rapid detectionof Salmonella,Streptococcus agalactiae and Staphylococcus aureus.Methods：Based on Histidine transport operon gene in Salmonella,STRA-Agl-23-1D genes in Streptococcus agalactiae and Heat-resistant nuclease nuc gene in Staphylococcus aureus,three pairs of primers were designed for PCR amplification and its reaction conditions were optimized.Results：Under the optimized conditions,the three fragments of 495 bp,360 bp and 279 bp were simultaneously amplified from three couple primers.The M-PCR assays for the simulation samples indicated that the detection limits were 1.9×102 cfu/mLl for Salmonella,2.0×102 cfu/mL for Streptococcus agalactiae and 2.9×102 cfu/mL for Staphylococcus aureus.The detection limit of DNA content of the three strains were 3.86、 25.5、 and 3.47pg,respectively.Conclusions：A simple,rapidand sensitive multiplex PCR detection method has been established,with good prospects.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2010年第6期173-179,共7页
Journal of Chinese Institute Of Food Science and Technology
基金
"十一五"国家科技支撑计划重点项目(2009BADB9B06)
北京市科技计划项目(D1011050460000)
