应用CODEHOP PCR快速检测水体中致病性弧菌  

Rapid detection of pathogenic Vibrios in water by CODEHOP PCR

作　　者：胡群[1] 邹春颖[1] 童淑梅[1] 马思杰[1] 

机构地区：[1]大榭出入境检验检疫局,浙江宁波315812

出　　处：《中国卫生检验杂志》2014年第16期2346-2348,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的建立可以快速检测不同致病性弧菌的CODEHOP PCR方法。方法以致病性弧菌共有的DNA旋转酶B亚单位氨基酸为目标,采用CODEHOP程序,设计一对引物,建立弧菌CODEHOP PCR检测方法,以8种不同致病性弧菌菌株及其他菌株为模板,评价方法的灵敏度、特异性,并对64份入境船舶压舱水进行致病弧菌检测。结果该方法特异性好,8种致病性弧菌标准菌株均获得特异性扩增条带,检测灵敏度可以达到3×103CFU/ml,从64份入境压舱水中检测到14份阳性样品,PCR产物经TA克隆后,测序分析确定为4种不同致病性弧菌。结论本研究所建立的方法可用于致病性弧菌的快速检测与种类鉴定。Objective To develop a method for fast detection of different pathogenic Vibrios by PCR with consensus degenerate hybridoligonucleotide primers（CODEHOPs）. Methods According to the amino acid sequence of DNA gyrase subunit B of dif- ferent pathogenic Vibrios, a pair of primers were designed by CODEHOP program, then a CODEHOP PCR method for detection of Vibrios was developed, and the specificity and sensitivity of the method were evaluated by using 8 different pathogenic Vibrios standard strains. Furthermore, the method was used to detect pathogenic Vibrios in 64 imported vessels＇ ballast water samples. Results The method showed better specificity, and specific amplification bands can be obtained in all the 8 pathogenic Vibrios standard strains. The detection sensitivity can reach 3 × 10 3 CFU/mh Fourteen samples were positive from 64 imported ballast water samples. PCR products, after TA cloning and sequencing analysis, were identified as 4 kinds of different pathogenic Vibri- os. Conclusion The method developed in this study can be used for rapid detection and identification of pathogenic Vibrios species

关 键 词：弧菌 检测 一致-简并杂合寡核苷酸引物 水体 

分 类 号：R378.3[医药卫生—病原生物学]