Abstract：Objective To investigate the HBV positive rate and the reasons of HBsAg miss detection in the HBsAg negative blood donors screened by ELISA , and to analyze the features of HBV genotypes and serotypes as well as the relationship between amino acid substitution of HBsAg S region and the ELISA miss detection. Methods A total of 31,184 HBsAg negative blood samples screened by ELISA were collected from November 2013 to October 2015 and tested for HBV DNA and HBV nucleus-related antigen (HBVNRAg). HBV gene sequence was then determined and analyzed in HBV DNA positive samples. Results All the 31,184 HBsAg negative blood samples were HBVNRAg negative. 82 samples were HBV DNA positive, with a detection rate of 0.26% (82/31,184). Among the 82 samples, 79 (79/82, 96.3%) had HBV genotype Band 3(3/82,3.7%) had HBV genotype C. All samples had serotype adw. S mutations were detected in 27 (27/82, 32.9%) samples,among which 21 amino acid mutation sites were located in 99a.a.-169a.a. at major hydrophilic region (MHR) of S protein, withthe major sites at 133 a.a.(7/82, 8.5%), 126 a.a (6/82, 7.3%), 161 a.a. (5/82, 6.1%), 134 a.a. (4/82, 4.8%) and 145 a.a. (2/82, 2.4%). 47 mutations were also detected in 21 amino acid sites (121, 122, 125, 128, 129, 131, 132, 140, 143, 150, 156, 157, 158, 159, 164, 166). Conclusions For blood donor screening, HBVNRAg detection is not significant. One of the main reasons of ELISA miss detection is the immune escape mutation of MHR located in 99-169 amino acid sites. Other reasons may be low blood HBsAg concentration and insensitive detection reagent. HBV DNA detection in blood screening can effectively reduce the miss detection rate by ELISA alone and significantly improve blood safety.
[1] 卢姗姗, 徐东平, 李进. HBV前S/S基因突变的研究进展[J]. 传染病信息, 2016, 29(2):171-173. [2] 陈秉宇, 沈健. HBsAg阴性献血者HBV-DNA检测的临床意义[J]. 中华医院感染学杂志, 2007, 17(10):1240-1241. [3] 叶贤林, 李活, 许晓绚,等. 核酸扩增技术在献血者血液HBV DNA、HCVRNA及HIV-1RNA筛查中的应用研究[J].中国输血杂志, 2010, 23(1):6-10. [4] 黄建国, 谢秀华, 欧阳玲,等. HBsAg阴性献血者人群HBV感染的检测和输血残余风险分析[J]. 检验医学与临床, 2011, 8(7):774-776. [5] 曾劲峰, 郑欣, 熊文,等. 核酸筛查后HBsAg-/NAT+/HBV DNA-献血者标本的确认试验[J]. 中国输血杂志, 2013, 26(12):1215-1218. [6] Torresi J, Earnest L, Deliyannis G, et al. Reduced antigenicity of the hepatitis B virus HBsAg protein arising as a consequence of sequence changes in the overlapping polymerase gene that are selected by lamivudine therapy[J].Virology, 2002, 293(2):305-313. [7] Echevarría JM, Avellón A. Hepatitis B virus genetic diversity[J].Med Virol, 2006, 78(Suppl 1):S36-S42. [8] Borroto-Esoda K, Miller MD, Arterburn S. Pooled analysis of amino acid changes in the HBV polymerase in patients from four major adefovir dipivoxil clinical trials[J].Hepatol, 2007, 47(4):492-498. [9] Yuan Q, Ge SX, Yan Q, et al.Establishment of a new combined enzyme immunoassay for detection of HBV prs1 and core antigens and the consistency with HBV DNA test.[J].Bing Du Xue Bao,2007,23(4):252-257. [10] Kimura T, Rokuhara A, Matsumoto A, et al. New enzyme immunoassay for detection of hepatitis B virus core antigen (HBcAg) and relation between levels of HBcAg and HBV DNA[J].J Clin Microbiol, 2003, 41(5):1901-1906. [11] Yuan Q,Ge SX,Xiong JH. A novel immunoassay for PreS1 and/or core-related antigens for detection of HBsAg variants[J].J Virol Methods, 2010,168(1-2):108-113. [12] 章述军, 田德英, 李晖,等.联合检测HBV前S1抗原和核心抗原的临床意义[J].中西医结合肝病杂志,2008,18(1):74-77. [13] 王林川,于燕,徐莉. 乙型肝炎病毒核酸相关抗原检测及其临床意义[J].国外医学医学地理分册,2011,32(4):261-264. [14] Huang YH,Huang HH.Core antigen expression is assoiated with hepatic necroinflammation in e antigen-negative chronic hepatitis B patients with low DNA loads[J].J Clin Vaccine Immunol, 2010,17(1):33-35. [15] 胡伟,胡孝彬,代琼.慢性乙型肝炎病毒感染者血清乙型肝炎病毒相关抗原检测的应用价值[J].实用医技杂志,2013,20(10):1055-1057. [16] Chinese Society of Hepatology, Chinese Meldical Association,Chinese Society of Infectious Diseases, Chinese Medical Association. Guideline on prevention and treatment of chronic hepatitis B in China (2005)[J]. Chin Med J (Engl),2007, 120(24):2159-2173. [17] Lok AS, McMahon BJ. Chronic hepatitis B: update 2009[J]. J Hepatol, 2009, 50(3):661-662. [18] Lok AS. Evolution of nucleoside/tide analogues for hepatitis B: Is the ideal drug here yet?[J]. J Hepatol, 2009, 51(2):416-418. [19] Sheldon J, Soriano V. Hepatitis B virus escape mutants induced by antiviral therapy[J]. J Antimicrob Chemoth, 2008, 61(4):766-768. [20] Villet S, Pichoud C, Billioud G, et al. Impact of hepatitis B virus rtA181V/T mutants on hepatitis B treatment failure[J].JHepatol, 2008, 48(5):747-755.
2017, Vol. 24 
