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目的探索适合于基层实验室开展的水中诺如病毒浓缩与应急检测的方法,评估钙离子法对水体中诺如病毒的富集效果及检测灵敏度,为水源性诺如病毒突发疫情的病原检测提供可靠的检测数据。方法将不同浓度的指示病毒(GⅡ型诺如病毒)加入到不同水样中,用钙离子法进行富集,用荧光定量PCR检测,根据富集前后的病毒浓度,评估方法的回收率;根据不同梯度的检测值,评估方法的灵敏度。同时制备不同的模拟水样进行荧光定量PCR检测。结果钙离子法的平均回收率为81.6%,检测灵敏度为模拟样本中病毒浓度为10 copy/μl。而10份不同模拟水样荧光定量PCR检测阳性9份。结论钙离子法浓缩水样中的诺如病毒,回收率与灵敏度均较高,操作简单;同时,荧光定量PCR检测方法具有快速、灵敏的优点,适用于基层实验室开展水体中诺如病毒的应急检测。
Objective To explore a suitable method for concentration and emergency detection of Norovirus in grass-roots laboratories and to evaluate the effect of Ca 2+ -concentration on the concentration of Norovirus in water and the detection sensitivity, and to detect the pathogen of waterborne Norovirus virus Provide reliable test data. Methods Different concentrations of indicator virus (Gnorovirus Ⅱ) were added into different water samples and enriched by calcium ion. The fluorescence quantitative PCR was used to detect the recovery rate of the method according to the concentration of virus before and after enrichment. The different gradient of the test value, evaluate the sensitivity of the method. At the same time, different simulated water samples were prepared for fluorescence quantitative PCR detection. Results The average recovery rate of calcium ion method was 81.6%. The detection sensitivity was 10 copy / μl in the simulated sample. And 10 different simulated water samples were detected by fluorescence quantitative PCR 9 positive. Conclusions The method of calcium ion method for the determination of norovirus in water sample has high recovery rate and sensitivity and simple operation. Meanwhile, the method of fluorescence quantitative PCR has the advantages of rapid and sensitive, and is suitable for the grassroots laboratory to carry out Norovirus Emergency detection.