霍乱弧菌的PCR方法检测及耐药性分析

目的建立PCR检测霍乱弧菌的快速方法,应用于水源、海产品霍乱弧菌快速检测;了解霍乱弧菌的耐药性。方法根据霍乱弧菌的肠毒素基因A亚单位(ctxA)的保守序列,设计引物,以副溶血弧菌和沙门菌为对照,建立PCR检测霍乱弧菌的快速方法,用于霍乱弧菌的快速检测;应用美国德灵公司生产的Walkway40微生物鉴定和药敏分析系统对70株霍乱弧菌进行耐药性检测。结果70株霍乱弧菌出现特异性荧光,其他菌不出现荧光,灵敏度高、特异性强,可在8h内作出诊断。霍乱弧菌对大部分抗生素敏感,对复方新诺明耐药率较高。结论PCR检测霍乱弧菌灵敏度高特异性强可用于水源、海产品霍乱弧菌快速检测;霍乱弧菌对大部分抗生素敏感。

The detection of vibrio cholerae with PCR methods and the analysis of the drug-resistance

Objective The study was conducted to develop PCR methods for the rapid detection of vibrio cholerae in water sources and seafood products and to get insight into the drug-resistance of the organism. Methods The primers were designed based on the conserved sequence of ctxA and PCR methods for the rapid detection of vibrio cholerae were developed with bibrio parahemolyticus and saimonella as control groups. The Microbe/Medicament Analyzing System (Walkway 40, DADE BEHRING INC, America) was used for the detection of drug-resistance of seventy strains of vibrio cholerae. Results Specific fluorescence was found in all the 70 strains of vibrio cholerae, but not in other bacteria, which indicated that the technique had high sensitivity and high specificity with diagnosis made within eight hours. It was found that the bacteria were sensitive to most of antibiotics but highly resistant to SMZ (Cotrimoxazole). Conclusion PCR methods are of high sensitivity and specificity in the detection of vibrio cholerae, applicable to the rapid detection of the bacteria in water sources and seafood products. Vibrio cholerae are highly sensitive to most of antibiotics.