Comparison of methods for the detection of hepatitis E virus in simulated water samples
10.3760/cma.j.cn112866-20231101-00048
- VernacularTitle:模拟水样中戊型肝炎病毒检测方法的比较
- Author:
Ruiting ZHANG
1
;
Qiuyuan WANG
;
Wenjiao YIN
;
Jingyuan CAO
;
Shengli BI
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 国家卫生健康委员会医学病毒和病毒病重点实验室，北京 102206
- Keywords:
Hepatitis E virus;
Concentration;
Pretreatment method;
Real time RT-PCR
- From:
Chinese Journal of Experimental and Clinical Virology
2024;38(1):93-98
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To compare the detection method of hepatitis E virus (HEV) in simulated water samples, and to provide a reference for the detection of HEV in water.Methods:HEV fecal suspension was added to tap water or distilled water simulated water samples, and pretreatment was carried out by electropositive filter-organic eluent elution method (Method 1) to compare the extraction effect of the three nucleic acid extraction kits, A, B, and C. The simulated water samples were pre-treated by Method 1, 2 (electropositive filter-direct lysis method), 3 (tangential-flow ultrafiltration membrane-organic eluent elution method), and 4 (tangential-flow ultrafiltration membrane-direct lysis method) for pretreatment, A kit for nucleic acid extraction, Real time RT-PCR method for detection and comparison of the recovery rate; comparison of the recovery rate of different concentrations of HEV in simulated water samples; comparing the inhibitory effects of inhibitors in tap water samples on real time RT-PCR; and detection of HEV in different batches of tap water specimens.Results:Kit A nucleic acid extraction was better; the recoveries of method 1, 2, 3 and 4 were 7.31%, 39.88%, 6.85% and 64.88%, respectively, which showed a statistically significant difference in the recoveries ( F=114.069, P<0.001). The recoveries of method 4 with the addition of high, medium and low concentrations of HEV were 65.26%, 42.76% and 32.79%, respectively. The inhibition of all four pre-treatment method was less than 75%, which meets the requirements of ISO (15216-2∶2019). Twenty tap water specimens were tested for HEV and the result were negative. Conclusions:This study showed that the two membranes better recovered in combination with direct lysis, respectively; Methods 4 had a higher recovery in the detection of HEV in small volumes of distilled or tap water, but it was limited by the volume of water samples, turbidity, and so on. Suitable method can be selected for different water quality and laboratory conditions.
