论文部分内容阅读
目的比较应用传统培养和PCR技术检测引起食物中毒的食品和患者肛拭子副溶血弧菌的方法。方法按照国家标准GB4789和有关规范采集引起食物中毒的食品和患者样品,采用传统培养方法分离鉴定病原菌,并结合分子生物学技术提取食品和患者样品的总DNA、PCR扩增病原菌特异性基因。结果 12份患者肛拭子样品中有8份样本分离培养出同一血清型O3:K6的副溶血弧菌,8份样本PCR扩增副溶血弧菌毒力基因结果也呈阳性;患者食用过的海鲜等18份冷冻食品采用传统培养方法均未分离到活的副溶血弧菌,但其中有2份食品PCR扩增结果呈阳性。结论本研究表明,患者的肛拭子用传统培养方法分离鉴定副溶血弧菌结果与PCR扩增结果一致;而冷冻的海鲜食品中副溶血弧菌也许进入了“活的非可培养(VBNC)”状态,难以通过分离培养方法检测出来,但用PCR技术能扩增出其存在的特异性基因,从而提高检测病原菌的敏感性。
Objective To compare the methods of traditional culture and PCR detection of food poisoning caused by food poisoning and Vibrio parahaemolyticus in patients with rectal swab. Methods Samples of food and patient causing food poisoning were collected according to the national standard GB4789 and relevant regulations. The pathogen was isolated and identified by traditional culture method. The total DNA of food samples and patient samples were extracted by molecular biology techniques, and the specific genes were amplified by PCR. Results Eight samples from 12 samples of anal swabs were isolated and cultured to produce Vibrio parahaemolyticus of the same serotype O3: K6, and the virulence genes of Vibrio parahaemolyticus from 8 samples were also positive. The edible 18 strains of frozen seafood such as seafood were not isolated by conventional culture methods of live Vibrio parahaemolyticus, but 2 of them showed positive PCR results. CONCLUSIONS: This study shows that the isolation and identification of Vibrio parahaemolyticus by the traditional culture method in patients with anal swabs is consistent with the results of PCR amplification. However, Vibrio parahaemolyticus may enter the “non-culturable VBNC ) ”State, it is difficult to detect by isolation and culture methods, but using PCR technology can amplify its specific genes, thereby increasing the sensitivity of detecting pathogens.