摘要
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 the suitable way of the norovirus concentration of water and emergency detection for basic laboratory and assess the enrichment and detection sensitivity of the calciumion method on norovirus in water and provide reliable testing data for the pathogen detection of waterborne norovirus＇ s outbreak. Methods Add different concentrations of indicator virus（ GⅡnorovirus） into different water samples and enrich the samples by calciumion method and detect it by quantitative PCR of fluorescence. Assess the rate of recovery of norovirus according to the different concentrations of virus after enrichment and assess the method＇s sensitivity according to the different test results. Meanwhile,prepare different simulated water samples and detect them with quantitative PCR of fluorescence. Results The recovery rate of calcium method was 81. 6%,the sensitivity was 10 copies per μl of virus concentration in simulated samples. And there are 9 positive samples in 10 different simulated water samples by PCR. Conclusion The recovery rate and sensitivity of norovirus in condensed water samples by calciumion are both high and its operation is simple; at the same time,quantitative PCR of fluorescence is a rapid and sensitive detection method,which is suitable for emergency detection for norovirus in water of basic laboratory.
出处
《中国卫生检验杂志》
CAS
2015年第7期977-979,共3页
Chinese Journal of Health Laboratory Technology
关键词
诺如病毒
水样富集
荧光定量PCR
应急检测
Norovirus sample enrichment
Realtime fluores-cence quantitative PCR
Emergency detection
