Comparison of four nucleic acid detection methods for hepatitis A virus
10.3760/cma.j.cn112866-20201214-00304
- VernacularTitle:甲型肝炎病毒四种核酸检测方法的比较
- Author:
Feng SHI
1
;
Jingyuan CAO
;
Feng QIU
;
Wenjiao YIN
;
Wenting ZHOU
;
Shengli BI
Author Information
1. 中国疾病预防控制中心病毒病预防控制所，北京 102206
- Keywords:
Hepatitis A virus(HAV);
Fluorescence quantitative Polymerase Chain Reaction;
droplet chip digital RT-PCR
- From:
Chinese Journal of Experimental and Clinical Virology
2021;35(2):213-217
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To compare the four nucleic acid detection method of hepatitis A virus.Methods:Using method A, B, and C real-time fluorescent quantitative RT-PCR(RT-qPCR)and method D droplet chip digital PCR(RT-dPCR)to detect the sensitivity of HAV plasmid and gradient dilution HAV vaccine respectively. Specific detection of related viral nucleic acid was performed. Methods A, B, and C were used to detect 40 artificially contaminated HAV oysters, commercially available oysters and serum samples, and HAV vaccine samples, and compare the detection rates. The recovery rates of method A and D on artificially contaminated oysters were compared with low concentration of HAV.Results:Both method A and B could detect HAV plasmids up to 10 copies/μL. In the detection of HAV vaccine with gradient dilution, the slope, R 2 value and amplification efficiency of method A, B, and C were all within the acceptable range (-3.446~-3.297, 0.991-0.998, -95.07%-101.051%). For 40 specimens from different sources, the positive detection rates of method A, B, and C were 50% (20/40), 47.5% (19/40), 55% (22/40), and the difference was not statistically significant ( χ2=0.467, P=0.792). Methods A and D have no significant difference in the detection sensitivity of gradient dilution vaccines. For the detection of artificially contaminated oysters with low concentration of HAV, the recovery rate of method D was higher than that of method A, but the difference was not statistically significant (F=0.294, P=0.642). Conclusions:There is no significant difference between method A, B, and C, which is more convenient and fast. When detecting low concentrations of HAV in food, Methods D had a slight advantage, but the detection cost is slightly higher. The detection method can be selected according to the actual situation.
