Comparison of enrichment and detection methods for hepatitis E virus in beverages represented by cola
10.3760/cma.j.cn112866-20241212-00175
- VernacularTitle:以可乐为代表的饮品中戊型肝炎病毒富集及检测方法比较
- Author:
Ruiting ZHANG
1
;
Qiuyuan WANG
1
;
Wenjiao YIN
1
;
Jingyuan CAO
1
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
- Publication Type:Journal Article
- Keywords:
Hepatitis E virus;
Cola;
Enrichment;
RT-qPCR;
RT-dPCR
- From:
Chinese Journal of Experimental and Clinical Virology
2025;39(1):122-127
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To compare enrichment and nucleic acid detection method for hepatitis E virus (HEV) in simulated cola samples.Methods:Cola samples experimentally contaminated with HEV were enriched using positively charged filter membrane-direct lysis (Method 1), tangential flow ultrafiltration membrane-direct lysis (Method 2), and Method 3 and 4 (with the addition of a PCR inhibitor removal step on the basis of Method 1 and 2, respectively), and were assayed by real-time fluorescence quantitative RT-PCR(RT-qPCR), and the recoveries and inhibition rates were compared. Digital RT-PCR(RT-dPCR) and RT-qPCR were applied to detect the recovery of HEV in different medium and low concentrations of experimentally contaminated cola samples; and the inhibition rate and sensitivity of HEV RNA detection in different matrices.Methods 3 was selected for virus enrichment of 8 commercially available cola specimens, RT-qPCR and RT-dPCR for HEV RNA detection.Results:The HEV recoveries of method 3 and 4 (10.44% and 10.16%) were higher than those of method 1 and 2 (4.89% and 0.32%), and the differences were statistically significant ( P<0.05). The inhibition rates of method 3 and 4 were smaller than the inhibition rates of method 1 and 2. The recoveries of HEV in medium concentration artificially contaminated cola samples by RT-qPCR and RT-dPCR were 17.04% and 16.28%, respectively, and for low concentration artificially contaminated cola samples were 6.91% and 4.65%, respectively, and the differences in recoveries between the two assays at the same concentration were not statistically significant ( P=0.260, P=0.107 ); Cola matrix inhibits the detection of both RT-qPCR and RT-dPCR assays. Eight commercially available cola specimens were negative for HEV. Conclusions:Detection of HEV in cola beverages can be done by positively charged filter membrane-direct lysis + inhibitor removal (method 3) or tangential flow ultrafiltration membrane-direct lysis + inhibitor removal (method 4) enrichment, followed by RT-dPCR or RT-qPCR, with a high recovery of virus detection.
