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当前位置: 首页 > 文章 > 应用PCR技术快速测定食品中单核细胞增生李斯特氏菌毒力 食品科学 2010,31 (23) 164-168
Position: Home > Articles > Rapid PCR Detection of Listeria monocytogenes virulence in Food FOOD SCIENCE 2010,31 (23) 164-168

应用PCR技术快速测定食品中单核细胞增生李斯特氏菌毒力

作  者：
于丰宇;李林;王红;王文斟;何源;刘晓朋;凌华
单  位：
西南大学食品科学学院;重庆市疾病预防控制中心
关键词：
单核细胞增生李斯特菌;毒力;基因;聚合酶链式反应
摘  要：
目的:采用PCR技术准确、快速测定单核细胞增生李斯特菌(单增李斯特氏菌)分离株的毒力基因,鉴别强毒株和弱毒株或无毒株,以有效地限制李斯特氏菌病的传播。方法:以单增李斯特氏菌相关毒力基因(hly、plcB、inlA、inlB、inlC、inlJ、prfA)设计引物,检测重庆市2007—2009年分离的40株单增李斯特氏菌分离株中的毒力基因携带率,并进行小鼠毒力实验。结果:40株分离株中有15株7种毒力基因检测结果均为阳性,6株hly基因阴性,4株plcB基因阴性,4株prfA基因阴性,5株inlA、inlB基因阴性,11株inlC基因阴性,9株inlJ基因阴性,1株inlA、inlB、inlC、inlJ基因均为阴性。4株分离株的毒力与标准菌株ATCC191161E相当,小鼠LD50在1.2×108～6.0×108CFU/mL之间,为强毒株。筛选出弱毒株09-132,LD50为1.7×1011CFU/mL。结论:重庆市存在发生李斯特菌食物中毒的潜在危险,内化素基因(inlA、inlB、inlC、inlJ)的缺失可能是导致菌株毒力降低的原因,而prfA和plcB基因与菌株毒力的相关性较小。
译  名：
Rapid PCR Detection of Listeria monocytogenes virulence in Food
作  者：
YU Feng-yu1,LI Lin1,WANG Hong2,WANG Wen-zhen2,HE Yuan2,LIU Xiao-peng2, LING Hua2 (1. College of Food Science, Southwest University, Chongqing 400715, China; 2. Chongqing Center for Disease Control and Prevention, Chongqing 400042, China)
关键词：
Listeria monocytogenes;virulence;gene;PCR
摘  要：
Objective: The virulence-associated genes of Listeria monocytogenes (L.m) isolates were detected by PCR amplification assay in order to provide experimental references for evaluating the virulence of L.m isolates and effectively limiting the transmission of L.m. Methods: Forty strains of L.m isolated in Chongqing from 2007 to 2009 were subjected to the PCR detection of seven virulence-associated genes (hly, plcB, inlA, inlB, inlC, inlJ, prfA) and toxicity test in mice. Results: The PCR assay revealed that 15 of the 40 L.m isolates possessed all the seven virulence-associated genes, 6 strains were hly-, 4 strains were plcB-, 4 strains were prfA- , 5 strains were inlA-, inlB-, 11 strains were inlC -, 9 strains were inlJ -, and 1 strain was negative for inlA, inlB, inlC and inlJ. Four isolates were defined as virulent strains, whose virulence was equivalent to the standard strain, ATCC 1961E with a mouse LD50 between 1.2 × 108 CFU/mL and 6.0 × 108 CFU/mL. The LD50 of the screened low virulent strain 09-132 was 1.7 ×1011 CFU/mL . Conclusions: There is a potential risk of Listeria food poisoning in Chongqing City, and the internalized gene (inlA, inlB, inlC, inlJ) loss may lead to virulence declination, while the prfA and plcB genes are less relevant to the virulent isolates.

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