Two HBV DNA+/HBsAg− blood donors identified by HBV NAT in Shenzhen, China

Background

In order to further improve blood safety, mini-pool (MP) nucleic acid testing (NAT) was implemented to screen samples negative for hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (anti-HCV), anti-human immunodeficiency virus (anti-HIV), syphilis (anti-Treponemal antibody) and with normal ALT.

Study design and methods

From August 2006 to February 2008, 41,301 donations were screened using commercial HIV/HCV RNA and HBV DNA Real-Time PCR NAT assays in pools of 8. Reactive pools were re-tested as individual samples using the appropriate screening test and confirmed using an alternate commercial NAT assay. Donors reactive on both NAT assays were considered ‘confirmed’ positive for the virus concerned and recalled for additional follow-up testing and counseling.

Results

Of the 41,301 samples screened, no HIV or HCV RNA-positive/seronegative donations were detected but two HBV DNA positive/HBsAg negative blood donors (Donors 1 and 2) were identified. Their respective hepatitis immunological markers were: Donor 1 - anti-HBc positive/anti-HBe positive/HBeAg negative/ALT normal and HBV DNA viral load of 112 IU/ml; Donor 2 - anti-HBc positive/anti-HBe negative/HBeAg negative/ALT normal and HBV DNA viral load 2750 IU/ml.

Conclusions

MP NAT identified two HBsAg negative donors with presumed occult infection but no HIV or HCV seronegative/NAT positive (yield) donors. The HBV yield rate of 1 in 20,650 (95%CI - 1 in 5663 to 1 in 75,303) is comparatively high, exceeds the predicted rate based on previous modeling for the population and demonstrates the incremental blood safety value of NAT in countries where HBV is highly epidemic. The low viral load of the two yield samples underscores the importance of optimizing the sensitivity of the HBV NAT assay selected for screening.     

血液HBV DNA全自动检测及基因分型分析

目的建立血液HBVDNA标本汇集、核酸提取、扩增及检测的全自动筛查模式,对阳性标本进行基因分型和血清学追踪检测,为血液HBVDNA自动化筛查提供科学依据。方法在酶联免疫吸附试验(ELISA)筛查血液基础上,应用STAR2000加样仪自编程序进行全自动血样汇集(24人份),在MPLC仪上全自动提取标本核酸,应用PCR方法在COBASAMPLICOR进行扩增和检测分析结果,用国际标准核酸质控品考评检出限量,对阳性标本进行基因分型并追踪血清转换过程。结果全自动汇集、全自动核酸提取和扩增及检测HBVDNA95%检出限量为38.9IU/ml,95%CI为(21323)。通过对16512个标本共688个汇集池分析,HBVDNA阳性8例,阳性率为0.049%。其中C型3人,B型2人,D型1人,2人未能确定基因型,6例阳性标本追踪发现,3例发生了血清转换现象。结论HBVDNA全自动核酸检测方法可应用于24人份血液混样核酸筛查。     

HBsAg阴性献血者人群HBV感染的检测和输血残余风险分析

目的 了解献血人群乙型肝炎病毒(HBV)感染状况和血液经酶免疫法(EIA)筛查乙型肝炎病毒表面抗原(HBsAg)后经血传播HBV感染的残余风险.方法 采用国产和进口两种EIA试剂对献血者血液进行HBsAg筛查,罗氏诊断COBAS Ampliscreens NAT血筛系统检测EIA检测合格标本中HBV DNA,对HBV DNA阳性标本进行半套式PCR检测,并对PCR扩增产物进行测序和病毒基因亚型分析.结果 共筛查1998～2008年的献血者232 305例,发现HBsAg阳性2 999例,阳性率为1.3%;对2002～2007年EIA检测合格的113 639例献血者血液标本进行NAT检测,检测出13份HBV DNA阳性、HBsAg阴性的献血者血液,HBV残余风险高达1.1/10 000.结论 EIA筛查后血液安全性有了很好的保障,经血传播HBV残余风险依然处于较高的水平,NAT应用对提高血液安全,降低输血传播HBV残余风险意义重大.     

HBsAg阳性者前S1抗原和HBV DNA检测的相关性探讨

目的 探讨HBsAg阳性者血清中HBV DNA及乙型肝炎病毒前S1抗原(PreS1Ag)和乙型肝炎e抗原(HBeAg)的关系及其临床应用价值.方法 对368例临床血清标本采用ELISA方法检测HBeAg和PreS1Ag,荧光定量PCR法检测HBV DNA.结果 HBeAg阳性组PreS1Ag阳性率为87.2%,HBV DNA阳性率为91.0%;HBeAg阴性组PreS1Ag阳性率为54.7%,HBV DNA阳性率为55.7%.患者血清PreS1Ag与HBV DNA结果无明显差异(P>0.05).结论 PreS1Ag与HBV DNA符合率较高,比HBeAg更能反映病毒复制情况,可作为一项新的病毒复制指标.     

The clinical significance of occult hepatitis B transfusion in Taiwan – a look‐back study

Objectives: A look‐back study was conducted to determine the clinical significance of occult hepatitis B virus (HBV) blood transfusion in an HBV hyperendemic area. Aim: To improve the blood transfusion safety. Background: Occult HBV is transmissible through blood transfusion in HBV‐naÏve recipients. However, its impact on recipients with prevalent HBV infection in HBV hyperendemic areas is unclear. Methods/Materials: In 2006, 12 occult HBV blood donors were found from 10 824 repository samples by nucleic acid testing. The 74 corresponding recipients were identified and their pre‐ and post‐transfusion clinical information was gathered, and the living recipients were recalled for follow‐up. From the available archival sera, the HBV DNA was examined and sub‐genomic sequences between paired donor and recipient were compared using polymerase chain reaction‐based assays. Results: Among the 74 recipients, 18 were still alive and 12 returned to our clinic. From the available serological profiles, 76% of recipients had ongoing or recovered HBV infection before transfusion. Only 24 recipients had available post‐transfusion serological profiles and none seroconverted to be hepatitis B surface antigen (HBsAg) positive. Moreover, except for the prior HBsAg carriers, the recipients' HBV DNA levels after transfusion were low (<20 IU/mL). One recipient had identical HBV surface gene sub‐genomic sequence (384 nucleotides) to his donor. After transfusion, no recipient developed post‐transfusion hepatitis (PTH) and the clinical outcome was good. Conclusion: In HBV hyperendemic areas, occult hepatitis B transfusion might not lead to HBsAg carriage or PTH. The risk of transfusion‐transmitted HBV infection was probably lower than that in non‐endemic areas because most recipients had already experienced HBV infection.     

2010-2013年中国南京地区无偿献血人群HBV、HCV、HIV感染情况分析

本研究在于了解中国南京地区无偿献血者HBV、HCV和HIV感染及分布情况,探讨本地区人群的传染病流行趋势,为献血宣传和招募工作提供指导意见.将2010.6.10-2013.6.9三年间南京地区无偿献血者199777人次的血液标本,采用ELISA法检测HBsAg、抗-HCV和抗-HIV,对三项阴性标本再采用NAT联合检测HBV-DNA、HCV-RNA和HIV-RNA,对检测结果进行统计分析,并将抗-HIV初筛反应性标本送南京市疾病预防控制中心确证.结果表明,3年中无偿献血者HBsAg、抗-HCV、抗-HIV感染率分别为0.45％,0.28％,0.11％;NAT呈反应性为0.07％;确证抗-HIV阳性32例,其中男性30例（6例为再次献血者）,女性2例;不确定3例,其中男性2例,女性1例.经统计学处理证实,初次献血者与重复献血者HBsAg、抗-HCV、抗-HIV检测不合格率差异有统计学意义.结论：2010.6.10-2013.6.9南京地区无偿献血人群中抗-HCV、抗-HIV阳性率呈逐年下降趋势,HBsAg、NAT检测不合格率随年龄段增加而增加;抗-HCV、抗-HIV检测不合格率随年龄段增加而降低,重复献血者检测不合格率远远低于初次献血者,抗-HIV检测女性酶免检测阳性率高于男性,但确认阳性率男性明显高于女性.因此应提高献血前的征询技巧,重视献血者的招募环节,重点加强初次献血者献血前的征询和评估,加强无偿献血知识的宣传,巩固和发展固定无偿献血者队伍,大力普及NAT检测技术应用于血液筛查,将有效地提高血液安全.     

乙型肝炎病毒前S1抗原与HBV DNA和HBV-M及肝功能的相关性探讨

目的探讨兰州地区乙型肝炎病毒(HBV)感染者血清中前S1抗原(Pre-S1Ag)与HBV DNA、HBV-M及肝功能之间的相关性。方法对905例HBV感染者(HBV感染组)及100例健康体检者(健康对照组)进行Pre-S1Ag、HBV-M、HBV DNA和肝功能检测。结果 905例标本中,Pre-S1Ag和HBV DNA阳性率分别为68.51%(620/905)和67.96%(615/905),差异均无统计学意义(χ2=30.064,P0.05);570例HBeAg阳性组中,HBV Pre-S1阳性率为85.08%(485/570),显著高于HBeAg阴性组的Pre-SAg1阳性率40.30%(135/335),差异有统计学意义(χ2=108.881,P0.01)。Pre-S1Ag阳性组与阴性组ALT、AST异常率分别为53.22%、25.96%和51.29%、32.98%,差异有统计学意义(χ2ALT=53.148,P0.001,χ2AST=66.635,P0.001)。结论 Pre-S1Ag是乙肝病毒感染与复制的可靠指标,与HBV-DNA阳性相关度高,是对HBeAg的重要补充和加强,可为指导临床治疗提供更及时可靠的实验依据。     

全血乙型肝炎病毒定量检测方法的初步研究

目的探讨聚合酶链反应(PCR)检测全血中乙型肝炎病毒(HBV)DNA含量及临床应用价值。方法取20μl血清裂解液提取HBVDNA、和取同样量的全血经不同裂解液提取血液中血清及白细胞内总HBVDNA,同时经PCR检测。对78份乙型肝炎表面抗原(HBsAg)阳性患者两种取材方法检测HBVDNA含量进行比较。结果检测HBVDNA的最低检测限为1×103拷贝/ml,29份乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)和抗乙型肝炎核心抗体(抗HBc)均为阳性患者全血检测HBVDNA阳性率为100%,血清阳性率为100%,P>0.05;37份HBsAg、抗乙型肝炎e抗体(抗HBe)和抗HBc均为阳性患者全血检测HBVDNA阳性率为97.30%,血清阳性率为48.65%,P<0.01;12份HBsAg和抗HBc均为阳性患者全血检测HBVDNA阳性率为91.67%,血清阳性率为25%,P<0.01;20份非乙型肝炎患者血清均为阴性。血清HBVDNA阳性多数是ALT升高患者,其拷贝数均小于全血测定的拷贝数。结论全血检测HBVDNA阳性率高,避免血液中白细胞内HBV漏检,它能准确地反应血液HBV含量,更好地为临床治疗效果作监测。     

Analysis of HBV infection after blood transfusion in Japan through investigation of a comprehensive donor specimen repository

BACKGROUND: To understand the risk of transfusion-transmitted viral infection, it is important to precisely assess cases of infection that follow transfusion. STUDY DESIGN AND METHODS: HBV infections noted after transfusion in 1997, 1998, and 1999 were analyzed. Transfusion in all these cases was performed before NAT was adopted for donor screening. To detect viral infection, PCR and serologic tests for HBV were performed retrospectively on all blood samples from implicated donors that had been stored in a frozen state after each donation. The concentration of HBV genome was measured in HBV-positive blood samples. RESULTS: One hundred three cases of HBV infection were analyzed; of these, only 16, including at least 10 infections due to window-period (HBsAg-positive by reverse particle hemagglutination assay) donations, were confirmed by further testing to be related to transfusion. The concentrations of HBV genome were very low in four blood samples (<50, 400, 500, and 800 genome equivalents/mL of plasma). CONCLUSIONS: The remaining risk of transfusion transmission of HBV infection before the adoption of NAT was mainly due to window-period donations, including one that was made before the HBV genome was detectable by PCR. However, it was determined that transfusion was not responsible in many cases for HBV infection after transfusion.     

三种方法四种试剂自动化检测HBV血清标志物结果的对比研究

目的评价三种方法四种试剂自动化检测HBV血清标志物结果的差异。方法选择A xSYM、A lise i、SYM-B io自动免疫分析仪分别采用三种方法四种试剂对636例筛选的血清标本进行HB sA g、A n ti-HB s、HB eA g、A n ti-HB e、A n ti-HB c等五项血清标志物检测,并以A xSYM结果为参照,对项目结果不符及项目结果不一致进行分析。结果对636例筛选的血清标本,A lise i KHB、A lise iM urex、SYM-B io与A xSYM比较,其总的项目结果不符率和总的项目结果不一致率分别为12.4%和3.5%,11.1%和2.6%,7.5%和8.6%,各项目结果不符率以A lise iKHB、A lise iM urex的A n ti-HB e和SYM-B io的HB eA g/A n ti-HB c最高,分别为5.0%,5.8%和2.2%;各项目结果不一致率以A lise i KHB、A lise iM urex的A n ti-HB s和SYM-B io的A n ti-HB e最高,分别为2.3%,1.6%和3.2%。结论仪器、方法、试剂不同可导致血清标志物结果的差异,各项目的报告单位、判断标准有待进一步规范,应建立完善的检测血清标志物量值溯源体系。     

PreS1、乙肝两对半与HBV—DNA检测相关性分析

[目的]探讨慢性乙型病毒肝炎病人乙肝两对半（HBV—M）、前S1（PreS1）及HBV—DNA检测的相关性。[方法]HBV—M和PreS1采用ELISA法检测，HBV—DNA采用Real—time PCR法检测。[结果]131例慢性乙肝病人的HBV—M检测中，HBsAg（＋）、HBeAg（＋）、抗-HBc（＋）（1．3．5阳性）俗称“大三阳”，HBsAg（＋）、抗-HBe（＋）、抗-HBc（＋）（1．4．5阳性）俗称“小三阳”，HBsAg（＋）、抗-HBc（＋）（1．5阳性）3种典型感染模式为126例，占96．2％；其中72例1．3．5阳性病例中PreS1和HBV—DNA阳性分别为60例和72例，阳性率分别为83．3％和86．1％，而42例1．4．5阳性病例中PreS1和HBV—DNA阳性分别为12例和28例，阳性率为28．6％和66．7％；在78例PreS1阳性病人中HBV—DNA阳性占68例，其阳性率为87．2％，48例PreS1抗原阴性的病人中HBV—DNA阳性为32例，其阳性率为66．7％。[结论]在慢性乙型肝炎患者HBV—M检测“大三阳”模式中PreS1与HBV—DNA均表现高阳性率，二者具有良好的相关性（P〈0．01）；而“小三阳”模式中PreS1阳性率较低，但HBV—DNA仍有较高的阳性率，表明在慢性乙肝患者“小三阳”模式中HBV—DNA检测比PreS1检测更有价值（P〈0．01）。值得注意的是在我们的观察中PreS1阳性和阴性病人均有较高的HBV—DNA阳性率，二者没有明显差异（P〉0．05）。因而，在临床诊断、治疗和疗效观察中HBV—DNA可作为检测“金标准”，而PreS1检测时PreS1阴性的病人也不应忽视。     

HBV DNA定量检测方法研究进展

HBVDNA定量检测对乙型肝炎的诊治十分有用。近年来 ,许多检测方法得到广泛应用 ,如AmplicorPCR、bDNA技术和定量杂交试验等 ,现就其检测原理及优缺点作一阐述     

兰州地区 HBsAg、HBeAg、HBeAb、HBcAb 与 HBV DNA 的相关分析及临床价值

目的：探讨兰州地区慢性乙型肝炎病毒（HBV）感染者血清乙型肝炎病毒脱氧核糖核酸（HBV DNA）载量与乙型肝炎病毒血清学标志物（HBV-M）HBsAg、HBeAg、HBeAb、HBcAb 的相关性。方法724例 HBV 感染者分别利用实时荧光定量PCR 法检测 HBV DNA 载量,运用双抗体夹心化学发光免疫分析法检测血清 HBsAg、HBeAg、HBeAb、HBcAb 水平。结果兰州地区慢性 HBV 感染者 HBsAg 水平与 HBV DNA 载量呈正相关关系（r＝0．342,P ＜0．05）,HBeAg 水平与 HBV DNA 载量呈正相关关系（r＝0．463,P ＜0．05）,HBeAb 水平与 HBV DNA 载量之间呈负相关关系（r ＝-0．227,P ＝0．001）,HBcAb 水平与HBV DNA 载量变化无相关性（r＝-0．062,P ＝0．366）。结论兰州地区慢性 HBV 感染者 HBV DNA 载量和 HBsAg、HBeAg均有明显的正相关关系,提示临床上应把 HBsAg、HBeAg 与 HBV DNA 结合起来观察,能更准确判断患者的传染性程度。     

乙型肝炎病毒感染者HBeAg模式与病毒DNA水平相关性研究

目的 探讨乙型肝炎病毒(HBV)感染者HBeAg模式与HBV DNA水平之间的关系.方法 运用荧光定量PCR(FQ-PCR)检测120例乙型肝炎患者血清中的HBV DNA,同时运用ELISA方法进行HBV血清标志物的检测,并对结果进行相关性探讨.结果 HBeAg阳性组血清HBV DNA检出率为87.9%(58/66),...     

乙肝病毒感染者前S1蛋白和HBV—DNA相关性分析

目的探讨乙型病毒感染者前S1蛋白和HBV—DNA检测的相关性及临床价值。方法采用酶联免疫吸附试验（ELISA）方法检测748例乙肝患者HBV—M和前S1蛋白,并与HBV—DNA做对比分析。结果在HBsAg／HBeAg／HBcAb均阳性的高复制组中,前S1蛋白阳性占86．9％,HBV—DNA阳性占98．2％;HBsAg／HBeAb／ABeAb均阳性的低复制组中,前S1蛋白阳性占31．1％,HBV—DNA阳性占416;HBsAb／HBeAh／HBcAb均阳性的康复组中,前S1蛋白和HBV—DNA均为阴性。前S1能够敏感地反映乙型肝炎病毒的复制情况,尤其可以反映HBeAg阴性的乙型肝炎患者是否有病毒复制。结论前S1能够敏感地反映乙型肝炎病毒的复制情况,尤其可以反映HBeAg阴性的乙型肝炎患者是否有病毒复制。     

乙型肝炎病毒血清标志物与HBV-DNA定量联合检测的意义

目的探讨乙型肝炎（下称乙肝）病毒DNA（HBV-DNA）与血清免疫标志物的关系。方法荧光定量聚合酶链反应法（FO-PCR）检测血清样本中HBV—DNA含量;酶联免疫吸附法（ELISA）检测HBV血清标志物。结果A组（大三阳）、B组（小三阳）、C组[乙肝表面抗原（HBsAg）、乙肝e抗原（HBeAg）阳性]、D组（HBsAg、抗-HBc阳性）阳性率分别为97．3％、57．8％、100％、54．3％,HBeAg阳性组和HBeAg阴性组HBV-DNA病毒载量差异有统计学意义（P〈0．05）。结论HBV—DNA与乙肝血清标志物乙肝两对半（HBV-M）联合检测,能更准确地反映HBV复制情况,更有助于临床疾病的诊治。     

三种自动化免疫分析系统测定HBV血清标志物结果比较

目的　探讨不同的自动化免疫分析系统检测乙肝病毒血清标志物结果的一致性。方法　采用AxSYM、Elecsys 2 0 1 0和ARCHITECTi2 0 0 0三种自动化免疫分析系统分别对 1 2 0例随机选择的住院患者血清标本进行乙肝表面抗原 (HBsAg)及其抗体 (抗 HBs)、乙肝核心抗体 (抗 HBc)、乙肝e抗原 (HBeAg)及其抗体 (抗 HBe)等五项血清标志物测定 ,并以AxSYM结果为参照 ,计算三种方法所获结果的符合率。结果　对 1 2 0例随机血清标本 ,Elecsys 2 0 1 0与AxSYM比较测定结果符合率分别为HBsAg 99.2 %、抗 HBs 85 .8%、抗 HBc 87.5 %、HBeAg 99.2 %、抗 HBe 90 .0 % ;ARCHITECTi2 0 0 0与AxSYM比较测定结果符合率分别为HBsAg 1 0 0 % ,抗 HBs96 .7%、抗 HBc 91 .7%、HBeAg1 0 0 %以及抗 HBe95 8%。Elecsys 2 0 1 0与AxSYM测定抗 HBs的相关系数r =0 .90 ,ARCHITECTi2 0 0 0与AxSYMr =0 .98。Elecsys 2 0 1 0与AxSYM测定HBV五项标志物模式符合率为 6 5 .8% ,ARCHITECTi2 0 0 0与AxSYM测定五项标志物模式符合率为 85 .0 %。结论　三种方法测定HBV结果有一定差异 ,主要存在于三项抗体标志物以及低浓度抗原标志物的测定 ,有待进一步标准化     

乙型肝炎病毒DNA与标本物含量及肝功能生化指标间关系探讨

目的探讨血清中乙型肝炎病毒(HBV)DNA含量与HBV标志物[乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)]及肝功能指标间的相关性。方法抽取南平市人民医院肝内科住院的200例HBV携带患者血清样本。采用荧光定量聚合酶链反应(PCR)检测血清样本中HBV DNA含量,采用酶联免疫吸附试验(ELISA)测定血清样本中HBV标志物含量,同时检测肝功能。3项检测指标使用同一患者标本,分析3项指标之间的相关性。结果经相关性分析,HBV DNA与肝功能指标的r在±0.2以内,差异无统计学意义(P>0.05);HBsAg、HBeAg含量与肝功能指标的r在±0.2以内,差异无统计学意义(P>0.05);HBsAg、HBeAg含量与HBVDNA含量的r分别为0.457、0.568,差异有统计学意义(P<0.01)。结论 HBsAg、HBeAg含量只反映HBV在血清中的复制情况,不能以HBsAg、HBeAg含量及病毒载量的大小作为判断肝功能受损程度的指标;要综合分析HBV DNA含量与HBV标志物定量检测结果及肝功能,这样才能更有效地对疾病进行评估。     

Residual risk of HIV,HCV and HBV in Canada

Background

Residual risk is estimated as the product of the incidence and the infectious window period, the time during which a blood donation could be infectious but the assay may not detect it. In 2011 nucleic acid multiplex testing (MPX) was implemented in 6 unit minipools (previously 24 unit minipools). MPX also included hepatitis B (HBV) NAT for the first time (complementing HBsAg screening) in addition to HIV-1 and hepatitis C (HCV) as before. We aimed to estimate window period risk-day equivalents for MPX, and the residual risk of viral infections in blood donations updated to reflect current incidence and testing.