学术
荧光定量PCR快速检测食品中沙门氏菌方法的建立及初步应用
钟伟军[1] 赵明秋[1] 邓中平[2] 陈金顶[1] 徐艳芳[1]
[1]华南农业大学兽医学院,广东广州510642 [2]中山大学达安基因股份公司,广东广州510665
摘 要:
根据编码鼠伤寒沙门氏菌肠毒素stn基因的核苷酸序列设计1对引物和荧光探针，通过对荧光定量PCR反应体系和反应条件的摸索，建立了检测沙门氏菌的核酸荧光定量PCR方法。对该方法的特异性与敏感性研究结果显示，该方法检测沙门氏菌结果均为阳性，而非沙门氏菌均为阴性；对带有沙门氏菌肠毒素stn基因的阳性质粒的检测敏感性为4个／μL。用该方法对人工污染沙门氏菌的鲜猪肉和鲜鸡蛋进行检测，当检样中沙门氏菌初始含菌量分别为1CFU／g（鲜猪肉）和1CFU／g（鲜鸡蛋），经过12h的增菌后，检测结果均为阳性。该方法具有简便、快速、特异性强、敏感性高等特点。此研究为食品中沙门氏菌快速检测试剂盒的研制打下了良好的基础。[著者文摘]
文章出处:
《中国预防兽医学报》-2008年30卷3期 -220-224页
栏目信息:
分 类 号:
文献标识码:
A
文章编号:
1008-0589（2008）03-0220-05
Establishment and preliminary application of real-time PCR method for detection of Salmonella in food
ZHONG Wei-jun, ZHAO Ming-qiu, DENG Zhong-ping, CHEN Jin-ding, XU Yan-fang （1. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China; 2. Gangdong Da＇an Gene Co., Ltd. of Sun Yat-sen University, Guangzhou 510665, China）
Abstract:
A real-time PCR assay for detection of nucleotide of Salmonella typhimurium was established using a pair of primers and a probe designed according to the published nucleotide sequence of stn gene that encodes the entertoxin protein of Salmonella typhimurium. The assay had a detection limit of 4 copies/μL （100 copy/25 μL） for positive plasmid, and 1 CFU/g and 1 CFU/g for artificially contaminated fresh, meat and eggs, respectively. This method was rapid, highly sensitive and specific, which could be developed into a commercial kit for rapid detection of Salmonella typhimurium contaminations in food.[著者文摘]
Key words:
Salmonella; real-time polymerase chain reaction; rapid detection
基金资助:
国家自然科学基金.广东省自然科学基金联合项目（No.U0631006）;广州市科技攻关计划项目（No.200422-E0231）;广州市农业科技招标项目（No.GK0401001）
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