Comparison of detection methods for hepatitis E virus in pig liver
10.3760/cma.j.cn112866-20240429-00074
- VernacularTitle:猪肝中戊型肝炎病毒检测方法的比较
- Author:
Qiuyuan WANG
1
;
Ruiting ZHANG
;
Wenjiao YIN
;
Jingyuan CAO
;
Xiaomei LI
;
Shengli BI
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 国家卫生健康委医学病毒和病毒病重点实验室，北京 102206
- Keywords:
Hepatitis E virus;
Pig liver;
Virus recovery;
Nucleic acid detection;
Genotyping
- From:
Chinese Journal of Experimental and Clinical Virology
2024;38(5):570-577
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To optimize and compare method for hepatitis E virus (HEV) nucleic acid detection from pig liver, and provide technical references for HEV detection in animal viscera specimens.Methods:Three methods (PBS homogenization treatment, proteinase K treatment, chloroform extraction method) were used to pretreat and extract viral nucleic acid form pig liver, which was artificially contaminated with HEV fecal suspensions, and HEV RT-qPCR was used to compare the HEV recovery rate and inhibition rate. The optimized HEV method was applied to commercially available pig liver specimens, and HEV genotyping was performed on positive specimens.Results:The HEV recovery rate of PBS homogenization treatment, proteinase K treatment and chloroform extraction method was 9.88%, 0.19% and 17.28%, respectively. The recovery rate of proteinase K treatment was less than 1%, and it was discarded; t-test was performed to compare recovery rates of the other two methods, which showed statistically significant differences ( t=26.801, P<0.001), the chloroform extraction method had a higher recovery rate. The inhibition rates of the three methods were all less than 75%, within the range of the ISO/TS 15216-2∶2019 standard. Among 192 commercially available pig liver specimens, 17 specimens were detected positive for HEV RNA, with a nucleic acid positive rate of 8.85%; five specimens were successfully genotyped for HEV, all of which were genotype 4. Conclusions:The virus recovery effect was good when chloroform extraction method was used for pig liver pretreatment; moreover, this method could detect HEV RNA from commercially available pig livers, which indicate that it can be used for virus detection in food.
