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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 rapid detection method for foodborne pathogens Vibrio parahaemolyticus and apply it to the on-site rapid detection of food-borne outbreaks of pathogens in grass-roots laboratories. Methods Totally 239 clinical samples and 152 food samples were detected by the method of constant temperature amplification - nucleic acid test strip, fluorescence quantitative PCR and typical biochemical screening method. Through the standard strain test, observe the sensitivity and specificity of the method. Results 239 samples of clinical patients with diarrhea and 152 food samples were detected by the method of thermostatic amplification-nucleic acid test strip, real-time fluorescence quantitative PCR and typical biochemical screening method. The positive rate of clinical samples was 11.72% respectively. The positive rates of food samples were 19.73%, 20.39% and 20.39% respectively. There was no significant difference in the positive rates of the three methods (P> 0.05). There was no significant difference between the three test methods for food testing and specific test results (P> 0.05), but the sensitivity test results were lower limit thermostatic amplification-nucleic acid test strip method and real-time fluorescence quantitative PCR method were 10- 5 dilution (bacteria volume 22 cfu / ml), a typical biochemical screening method for the 10-4 dilution (strain 186 cfu / ml). Conclusion The method of thermostatic amplification - nucleic acid test strip has the advantages of short detection time, high sensitivity, strong specificity and simple instrument, which can be applied to the rapid detection in the field laboratory.