摘要
目的建立食源性病原菌副溶血性弧菌的快速检测方法,应用于基层实验室食源性暴发疫情病原的现场快速检测。方法采用恒温扩增-核酸试纸条方法、荧光定量PCR和典型生化联合筛检法对239份临床样本、152份食品样本进行平行检测比较。通过标准菌株试验,观察方法的敏感性与特异性。结果用恒温扩增-核酸试纸条法、实时荧光定量PCR法、典型生化联合筛检法同时对239份临床患者腹泻样本和152份食品样本进行检测,结果临床样本的阳性检出率均为11.72%,食品样本的阳性率分别为19.73%、20.39%、20.39%,3种方法的阳性率差异无统计学意义(P>0.05)。3种检测方法的食品检测、特异性试验结果差异均无统计学意义(P>0.05),但敏感性试验结果检测下限恒温扩增-核酸试纸条法、实时荧光定量PCR法均为10-5稀释度(菌量22 cfu/ml),典型生化联合筛检法为10-4稀释度(菌量186 cfu/ml)。结论恒温扩增-核酸试纸条方法具有检测时间短、灵敏度高、特异性强、仪器简单等优点,可应用于基层实验室的现场快速检测。
Objective To establish a method for the rapid detection of Vibrio parahaemolyticus,applied to the primary laboratory for screening pathogen in food borne disease outbreak. Methods Using isothermal amplification DNA test strip method,fluorescence quantitative PCR and typical biochemical combined screening method,239 clinical samples and 152 food samples were detected and compared. Through the parallel standard strain test,sensitivity and specificity were analyzed. Results 239 clinical specimens and 152 food samples were tested by the combined detection of the above 3 methods. The positive rate of clinical specimens was 11. 72%,and that of the food samples were 19. 73%,20. 39%,20. 39%,respectively,and the differences of positive rate among the 3 methods were not statistically significant（ P 〉0. 05）. The differences in food test and specificity test among the 3 methods had no statistical significance（ P 〉0. 05）,but in sensitivity,the lowest limit of isothermal amplification DNA test strip method and detection by real time fluorescent quantitative PCR was 10-5 dilution（ amount of bacterium contamination was 22 cfu/ml）,while it in typical biochemical combined screening method was 10-4 dilution（ amount of bacterium contamination was 186 cfu/ml）. Conclusion Isothermal amplification DNA test strip method has short detection time,high sensitivity,strong specificity,simple requirement of instrument,which can be applied to the primary laboratory in field fast measurement.
出处
《中国卫生检验杂志》
CAS
2017年第21期3089-3091,共3页
Chinese Journal of Health Laboratory Technology
关键词
食源性病原菌
副溶血性弧菌
恒温扩增
核酸试纸条
快速检测
Food borne pathogenic bacteria
Vibrio parahaemolyticus
Constant temperature amplification
Nuclein test strip
Fast measurement
