单人份核酸检测在血液乙型肝炎病毒筛查中的应用

目的 通过对血液标本的核酸和血清学的检测(NAT)结果进行比较分析,从而探讨核酸检测在血液病毒筛查中的作用.方法 对血液标本进行血清学和核酸检测.使用诺华诊断血液筛查系统对标本进行单人份核酸检测.如标本核酸检测为阳性,则需要对标本进行鉴别;鉴别结果为HBV DNA标本,采用电化学发光法进一步检测乙型肝炎血清标志物五项.结果 10 127例血液标本中检测出NAT(+)标本30例,其中NAT(+)、ELISA(-)的标本12例,ELISA漏检率为1.18‰,鉴别后有6例标本为HBV DNA(+)、ELISA(-),乙型肝炎血清标志物五项为全阴性或抗-HBc(+),未检出HIV RNA或HCV RNA;另有7例标本为NAT(-)、ELISA双试剂(+),其中3例为HBsAg(+),4例为抗-HCV(+).结论 核酸检测可以有效降低酶联免疫法漏检造成的输血风险,但其也存在漏检的情况,因此核酸检测和血清学检测相结合可作为血液筛查检测中的重要手段.     

合肥地区无偿献血者HBV感染调查及输血残余风险分析

目的了解合肥地区无偿献血人群HBV感染状况和经ELISA法筛查HBsAg后经血传播HBV的残余风险,为选择合适的血液筛查策略提供科学依据。方法采用两种ELISA试剂对献血者HBsAg进行筛查,同时用一种核酸检测系统检测标本中HBV DNA,HBsAg阴性HBV DNA阳性标本进行乙型肝炎血清标志物检测。结果共筛查献血者44 2567例,HBsAg阳性1 894例,阳性率0.428%;对68 662份标本进行核酸扩增检测,检测出45例HBsAg阴性HBV DNA阳性标本,HBV输血残余风险为0.066%。结论现有的ELISA检测体系存在输血传播HBV的风险,增加核酸检测能降低HBV输血残余风险。     

乌鲁木齐地区无偿献血者HBV核酸检测结果分析

目的了解乌鲁木齐地区无偿献血人群乙型肝炎病毒(HBV)感染的状况和无偿献血者的血液经酶免疫检测法(ELISA)筛查乙型肝炎病毒表面抗原(HBsAg)的感染状况,探讨采用核酸扩增技术(NAT)对血液进行筛查的可行性,以进一步改进献血者筛查方法,降低输血传播疾病残余风险。方法采用2种不同试剂厂家的ELISA试剂对无偿献血者血液进行HBsAg筛查,采用核酸扩增检测(NAT)血筛系统检测无偿献血者血的HBV DNA;并对ELISA检测阴性,HBV DNA阳性标本进行确认和追踪分析,以确定血清学感染状况。结果共筛查2011年3～10月乌鲁木齐地区无偿献血者14 696名,HBsAg阳性率为0.52%(76/14 696);HBV DNA阳性率0.22%(32/14696);检出2例HBV DNA阳性标本,经过追踪分析,第1例发生了血清学转换,为"窗口期"感染,第2例无血清学转换,但乙肝核心抗体持续阳性,为隐匿性乙型肝炎。结论 ELISA检测后血液安全性有了很好的保障,但是依然存在输血传播疾病残余风险,NAT检测可降低输血残余风险,从而提高输血安全。     

核酸检测与酶免检测平行筛查血液在基层血站的应用效果评价

目的探讨核酸检测与酶免检测平行筛查血液在基层血站的应用效果评价。方法选取2016年1月-2017年7月本基层血站无偿献血者标本34680例,采用核酸检测与酶免检测两种检测方法平行筛查,对酶免检测阴性的标本分别采用核酸检测NAT进行HBV DNA/HIV RNA/HCV RNA的检测。将核酸检测阳性标本进行分项定量检测和乙肝血清学补充试验,分析检测结果。结果经ELISA筛查HBsAg、抗-HCV和抗-HIV-1/2无反应性及仅1种试剂为反应性的标本共32456例,经核酸NAT检测阳性标本72例,阳性检出率为0.21%,ELISA筛查HB-sAg,抗-HBs,抗-HBe,HBeAg,抗-HBc,最终确认阳性标本48例,确认阳性率为66.67%(P0.05);72例NAT检测阳性标本进行定量检测,其中定量检测HBV DNA阳性36例,阳性率为75.00%(36/48),未检测出HIV RNA阳性和HCV RNA阳性。结论核酸检测与酶免检测平行筛查,两种手段相互补充,可使输血传播疾病残余风险极大得到降低,预防经输血传播的病毒性疾病,从可有效地保障血液安全。     

无偿献血者核酸检测实施效果评价

目的 通过分析无偿献血者全血标本的核酸检测(NAT)和血清学检测的结果,探讨NAT在筛查乙型肝炎病毒(HBV),丙型肝炎病毒(HCV)和人类免疫缺陷病毒(HIV)“窗口期”感染者或者隐匿性肝炎的患者中的作用.方法 选择2011年7月至12月于佛山市中心血站采集的无偿献血者全血样本13 126份作为研究对象.对无偿献血者全血标本同时进行血清学检测和NAT.使用诺华诊断血液筛查系统对标本进行单人份NAT.对于NAT联检和鉴别试验阳性而血清学试验阴性的献血者,进一步检测乙型肝炎血清标志物5项指标.结果 13 126份标本中,NAT联检试验阳性为128例(0.98％),其中95例为酶联免疫吸附试验(ELISA)双试剂阳性,4例为ELISA单试剂阳性,29例为ELISA阴性.对NAT联检试验和鉴别试验结果皆呈阳性的8例献血者做进一步追踪,6例行乙型肝炎血清标志物5项指标检测的结果显示,1例抗-HBs和抗-HBc呈阳性,3例仅抗-HBc呈阳性,2例呈全阴性.结论 NAT作为一种检测原理完全与ELISA不同的方法,能够在一定程度上弥补ELISA方法的局限性,有效缩短“窗口期”.NAT与ELISA两种检测方法能够互为补充,共同降低经输血途径传播病毒的风险.     

南宁地区献血者HBV感染的检测及输血残余风险评估

目的了解南宁地区无偿献血者HBV感染状况及血液在经过常规病毒血清学筛查后经血传播HBV病毒的残余风险,探讨本地区开展血液核酸筛查的意义。方法分别应用EIA法和速率法对2010年11月2日～2011年12月31日经快速法HBsAg和速率法ALT初筛合格的70 428份献血者血液标本进行常规病毒血清学筛查;应用罗氏Cobas S201核酸检测系统对68 716份经常规病毒血清学筛查合格的血液标本进行6样本混合法的HBV-HCV-HIV检测,并对阳性混样池进行拆分检测及对拆分检测阳性的标本送鉴别部门进一步分项鉴别确证。结果常规病毒血清学筛查中,HBsAg阳性标本有309份,阳性率为0.43%;核酸检测中,HBV DNA阳性标本有83份,阳性率为0.12%(1/828)。结论南宁地区常规病毒血清学筛查合格的献血者血液经血传播HBV的残余风险仍然处于较高的水平,核酸检测(NAT)的应用对提高血液安全,降低输血传播HBV残余风险的意义重大。     

无偿献血者血液HBV核酸筛查研究

目的 评估大连地区HBV-DNA检测对于输血安全的重要性.方法 对无偿献血者的血液标本进行血清学检测(HBsAg、HCV抗体、HIV抗原/抗体、梅毒特异性抗体、谷丙转氨酶和血型),同时进行HBV、HCV、HIV的三联检核酸检测(NAT);对ELISA(-)NAT(+)的标本采用电化学发光法进行HBV血清标志物补充试验,同时对献血者进行追踪检测.结果 22416份无偿献血者样本发现35例ELISA(+)NAT(+)、3例ELISA(+)NAT(-)、12例ELISA(-)NAT(+).跟踪5位ELISA(-)NAT(+)的献血者,1例可能是免疫“窗口期”;其余4例可能是OBI.结论 核酸血液筛查可大大提高血液安全性,对提高HBsAg阴性血液标本中HBV感染检出率具有重要价值;但核酸与血清学二者检测结果存在不一致,二者对于降低输血传播HBV.     

番禺地区无偿献血者酶免阴性样本的核酸检测结果分析

目的通过对酶联免疫吸附实验（ELISA）反应阴性样本的核酸检测技术（NAT）的筛查结果进行分析,探讨核酸检测技术在血液筛查中的应用价值。方法先采用ELISA试剂分别对献血者标本进行HBsAg、抗-HCV和抗-HIV筛查,筛查结果阴性的样本,再用罗氏诊断cobas s 201系统进行HIV、HCV和HBV三项联合NAT检测,并对NAT联检阳性的标本进行分项确证实验。结果在8 147例经ELISA筛查阴性血液标本中检出6例NAT阳性标本,经分项确证6例均为HBV DNA阳性,阳性检出率约为0.074%。结论 ELISA筛查阴性样本中仍存在一定的HBV DNA阳性结果,采用ELISA联合NAT筛查策略,能有效降低输血感染乙肝病毒的风险。     

核酸检测技术在南昌地区无偿献血血液筛查中的应用

目的病毒核酸检测技术(Nucleic Acid Technology,NAT)应用于献血者血液筛查,探讨缩短病毒检测"窗口期"的检测方法,保障临床输血安全。方法将2011年9月至2012年5月我站14203例无偿献血标本经血清学筛查合格的标本进行HBV、HCV和HIV三项联合NAT检测,并对NAT筛查阳性标本做确证试验。结果 HBsAg、抗HCV、抗HIV ELISA检测均为阴性的血液标本13843例,NAT共检出阳性25例,阳性率为0.18%。其中HIV-DNA检出1例,HBV-DNA检出24例。结论核酸检测技术应用于献血者血液筛查中,有效地缩短了病毒检出"窗口期",有力地保障了临床输血安全。     

合肥地区无偿献血人群HIV的检测及流行病学研究

目的了解合肥地区无偿献血人群HIV感染状况和流行病特点,为献血者招募和血液检测方案建立提供依据。方法采用两种血清学试剂对献血者进行筛查,2012年开始同时用核酸检测系统检测标本中HRV RNA,血清学阳性的标本送到市疾控中心做确认。结果血清学共筛查献血者495 279例,确认阳性44例,阳性率0.9;NAT检测标本112 940,确认阳性20例,全部为血清学双试剂阳性标本。结论合肥地区无偿献血人群HIV感染呈逐年上升趋势,必须采取有效措施保证血液安全。     

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.     

HBV DNA PCR检测在HBsAg阴性献血人群中的应用

目的 探讨HBsAg阴性献血者HBV DNA榆测的应用价值,评估核酸检测的必要性.方法 采用PCR检测HBsAg阴性献血者HBV DNA.采用8人份混合血样测定,超离心浓缩病毒,磁珠法提取病毒核酸.如HBV DNA为阳性,则进一步检测乙型肝炎病毒血清标志物5项.结果 HBVDNA检测限量为25 U/ml,23 225份标本中有4份为HBV DNA阳性,检出率为0.17‰.进一步检测其他HBV感染的血清学指标,发现这4份标本中有2份为抗HBe和抗HBc阳性,1份为抗HBc阳性,1份为抗HBs、抗HBc阳性.对HBV DNA的定量测定表明,其含量在50～200 U/ml.结论 现行的2次酶联免疫技术的血液筛查存在HBV漏检,有必要在现有的血液筛查模式中增加抗HBc检测,或增加病毒核酸筛查.     

Two cases of transfusion‐transmitted hepatitis B virus (HBV) infection in a low‐endemic country before implementation of HBV nucleic acid testing

BACKGROUND: The risk of hepatitis B virus (HBV) transmission by transfusion is higher than that of other blood‐borne viruses. In France, before the introduction of HBV nucleic acid testing (NAT) in 2010, blood donations were tested for hepatitis B surface antigen (HBsAg) and antibodies against hepatitis B core antigen, and the residual risk of HBV transfusion related to preseroconversion acute phase was estimated at 0.54 per million donations. The additional value of the implementation of a highly sensitive HBV NAT to prevent such transmissions is discussed. STUDY DESIGN AND METHODS: Two lookback investigations based on HBV seroconversion of repeat donors were performed. Donors and recipients were followed up in multiple samples that were tested for HBV serologic and molecular markers. RESULTS: The recipients have shown posttransfusion HBsAg seroconversion. The archived samples from the implicated donations were positive for HBV DNA at extremely low viral load in both cases. HBV isolates from donors and recipients of each case were organized in the same cluster with 100% identities into Genotypes A2 and B4, respectively. One recipient spontaneously recovered from infection while the second was successfully treated. CONCLUSION: The present cases highlight the importance of introducing highly sensitive HBV NAT to prevent transmission. Moreover, the lookback studies based on appropriate molecular and serologic investigations of patients transfused with previous donations from newly identified HBV‐infected repeat donors offer the opportunity to treat a recently infected recipient.     

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.     

无锡地区无偿献血者血液核酸筛查初步分析

目的初步分析罗氏MPX V1.0试剂在本地区献血者筛查中的应用情况。方法在罗氏Cobas核酸检测平台上,使用MPX V1.0试剂对9 418人次HBsAg、抗-HCV、抗-HIV、TP ELISA阴性的无偿献血者血液标本进行核酸定性检测,并对NAT阳性样本进行确认试验和电化学发光检测。结果 9 418人次标本共发现6个核酸检测阳性样本。对此类标本的部分献血者进行追踪,其中发现1例低CT值（MPX V1.0 16.4）样本,通过确认试验发现其为HCV感染,进行追踪检测证实该标本为HCV感染窗口期。结论应用罗氏MPX V1.0试剂对ELISA检测阴性的献血者血液标本进行筛查后,能进一步筛查出HBV、HCV或HIV核酸阳性的标本,从而阻止将该类血液提供到临床,进一步降低经血传播疾病的发生,提高血液质量,保证输血安全。     

上海地区无偿献血者乙肝病毒核酸检测分析

目的了解无偿献血者乙肝病毒核酸筛查(NAT)阳性人群特点,为血液安全策略提供参考。方法无偿献血者血液经Murex和科华HBsAg ELISA试剂检测,结果为阴性的血液使用cobas TaqScreen MPX试剂进行HBV DNA,HCV RNA,HIV RNA 3项联合核酸检测。对于MPX反应性标本,使用COBAS AmpliPrep/TaqMan进行核酸鉴别试验,同时使用罗氏ECL电化学发光检测系统进行乙肝补充血清学试验。结果 2011年11月1日～2012年1月31日3个月共有献血者86 375人(次),其中有63 351人(次)为初次献血者,HBsAg反应性为1.04%,23 024人(次)为重复献血者,HBsAg反应性为0.46%,两者差异有统计学意义(χ2=63.63,P0.05)。84 990份HBsAg、抗-HCV、抗-HIV1/2阴性血液进行MPX核酸检测,共发现52例(0.060%)HBV DNA阳性,均为低拷贝,含量为(20～200)IU/ml间,其中32例(0.051%)来自初次献血者,20例(0.087%)来自重复献血者,两者比例差异无统计学意义(χ2=3.65,P0.05),没有发现HCV RNA与HIV RNA阳性。结论重复献血者HBsAg反应性比率低于初次献血者;HBsAg阴性献血者HBV DNA阳性率为0.060%,重复献血者HBV DNA阳性率与初次献血者比较,两者差异无统计学意义;开展HBV核酸检测能够进一步保障血液安全。     

Efficacy of individual nucleic acid amplification testing in reducing the risk of transfusion‐transmitted hepatitis B virus infection in Switzerland,a low‐endemic region

BACKGROUND: The risk of transfusion‐transmitted hepatitis B virus (HBV) in Switzerland by testing blood donors for hepatitis B surface antigen (HBsAg) alone has been historically estimated at 1:160,000 transfusions. The Swiss health authorities decided not to introduce mandatory antibody to hepatitis B core antigen (anti‐HBc) testing but to evaluate the investigation of HBV nucleic acid testing (NAT). STUDY DESIGN AND METHODS: Between June 2007 and February 2009, a total of 306,000 donations were screened routinely for HBsAg and HBV DNA by triplex individual‐donation (ID)‐NAT (Ultrio assay on Tigris system, Gen‐Probe/Novartis Diagnostics). ID‐NAT repeatedly reactive donors were further characterized for HBV serologic markers and viral load by quantitative polymerase chain reaction. The relative sensitivity of screening for HBsAg, anti‐HBc, and HBV DNA was assessed. The residual HBV transmission risk of NAT with or without anti‐HBc and HBsAg was retrospectively estimated in a mathematical model. RESULTS: From the 306,000 blood donations, 31 were repeatedly Ultrio test reactive and confirmed HBV infected, of which 24 (77%) and 27 (87%) were HBsAg and anti‐HBc positive, respectively. Seven HBV‐NAT yields were identified (1:44,000), two pre‐HBsAg window period (WP) donations (1:153,000) and five occult HBV infections (1:61,000). Introduction of ID‐NAT reduced the risk of HBV WP transmission in repeat donors from 1:95,000 to 1:296,000. CONCLUSIONS: Triplex NAT screening reduced the HBV WP transmission risk approximately threefold. NAT alone was more efficacious than the combined use of HBsAg and anti‐HBc. The data from this study led to the decision to introduce sensitive HBV‐NAT screening in Switzerland. Our findings may be useful in designing more efficient and cost‐effective HBV screening strategies in low‐prevalence countries.