Application of nested PCR in detection of occult hepatitis B virus infection blood donors
10.3760/cma.j.cn112866-20221130-00248
- VernacularTitle:巢式PCR在献血者隐匿性乙肝病毒感染检测中的应用
- Author:
Hao WANG
1
;
Kewang SUN
;
Chao LIU
;
Xinping ZHANG
;
Xiangyan HUANG
Author Information
1. 中国人民解放军联勤保障部队第九六〇医院输血医学科（解放军济南血液中心），济南 250031
- Keywords:
Nested PCR;
Nucleic acid test;
Blood donors;
Occult hepatitis B virus infection
- From:
Chinese Journal of Experimental and Clinical Virology
2023;37(4):410-413
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the application of nested PCR in the detection of occult hepatitis B virus infection (OBI) in blood donors, and improve the detection rate of OBI in blood screening before blood transfusion.Methods:From July 2021 to August 2022, 37 blood donors who donated blood in our center were found to have HBsAg- and nucleic acid testing (NAT)+ by serological tests and nucleic acid amplification tests. The CT value of nucleic acid amplification test was recorded, and the HBV DNA was quantitatively detected by fluorescent quantitative PCR; Conventional PCR primers and nested PCR primers were designed to amplify HBV S gene and pre-S gene using conventional PCR and nested PCR respectively, and gene sequencing was performed for the amplification result.Results:Among the 37 HBsAg- and NAT+ blood donors, 33 nucleic acid detection CT values were in the gray area, and the fluorescent quantitative PCR detection of HBV DNA value was negatively correlated with the nucleic acid detection CT value; 37 cases of HBsAg-, NAT+ blood donors could not be amplified by conventional PCR. After using nested PCR, 34 cases could detect the S gene of HBV virus, and 28 cases could detect the pre S gene of HBV virus; The amplified bands were all HBV genome bands by gene sequencing.Conclusions:The plasma HBV viral load of HBsAg- and NAT+ blood donors was very low; nested PCR has high sensitivity, which can improve the detection rate of HBV S gene and pre-S gene, and has certain application value for the pre transfusion screening and follow-up research of OBI.
