摘要
目的:建立1种快速检测鸡肉中布氏弓形菌、嗜低温弓形菌和斯氏弓形菌的多重PCR方法。方法:根据3种弓形菌的rpoB基因设计了4条引物,进行多重PCR扩增,测试PCR体系的特异性和灵敏度,并应用于鸡肉样品的检测。结果:对布氏弓形菌、嗜低温弓形菌和斯氏弓形菌的基因组DNA能够特异性扩增出373,149和236 bp大小的目的条带,检测灵敏度分别为1,10,10 pg。16份鸡肉样品中,7份样品被检出布氏弓形菌,3份样品被检出嗜低温弓形菌,1份样品被检出斯氏弓形菌,与常规分离培养法检测的符合率分别为100%,87.5%和100%。结论:建立的多重PCR法具有操作简便,检测周期短,特异性强和灵敏度高的特点,能够实现对鸡肉中布氏弓形菌、嗜低温弓形菌和斯氏弓形菌的同时快速检测和监控。
Objective： A multiplex polymerase chain reaction （PCR） assay was developed for rapid detection of Ar- cobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii in chicken products. Methods： Four primers targeting rpoB genes were designed and the specificity and sensitivity of the multiplex PCR were determined. Then, A. butzleri, A. cryaerophilus and A. skirrowii isolated from chicken products were investigated using this PCR method. Results： The rpoB genes of A. butzleri, A. cryaerophilus and A. skirrowii were successful in specifically amplifying 373, 149 and 236 bp fragments, and these fragments could be detected at the minimum DNA weights of 1, 10, 10 pg, respectively, that 7, 3 and 1 of 16 chicken products were positive for A. butzleri, A. cryaerophilus and A. skirrowii, the coincidence rate with plate-culture method were 100%, 87.5% and 100%, respectively. Conclusion： The multiplex PCR assay is a sim- ple, rapid, sensitive and specific technique for detecting A. butzleri, A. cryaerophilus and A. skirrowii, and could be used for the simultaneous identification of them isolated from chicken products.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2014年第8期227-232,共6页
Journal of Chinese Institute Of Food Science and Technology
基金
广东省自然科学基金项目(S2013040013491)
关键词
多重PCR
布氏弓形菌
嗜低温弓形菌
斯氏弓形菌
Multiplex PCR
A rcobacter butzler
A rcobacter cryaerophilus
A rcobacter skirrowii
