核酸扩增技术在献血者血液HBV DNA、HCV RNA及HIV-1 RNA筛查中的应用研究

目的探讨在我国无偿献血者的血液筛查中引进核酸扩增技术(NAT)的必要性,了解献血者血清学病毒标志物检测阴性、NAT检测阳性的感染状况。方法应用Roche PCR、PCR-微流芯片、实时荧光PCR方法对深圳市131 174人(次)血清学检测病毒标志物阴性的献血者进行HBV DNA、HCV RNA和HIV-1 RNA检测,对NAT阳性献血者追踪检测并做定量分析。结果HBV DNA阳性22例,阳性率为1/5 962,其中15例为抗-HBc阳性,阳性率为1/8 745;HCV RNA阳性1例,阳性率1/131 174;HIV-1 RNA未检出阳性。对14名HBV DNA阳性者的追踪发现,8人发生了血清转换现象。结论采用高灵敏度的NAT筛查献血者血液中的HBV和HCV,有助于提高输血及血液安全。     

核酸检测技术应用于血清学筛查合格的献血者标本检测

核酸检测(nucleic acid test,NAT)是一种新兴的血液传染病检测方法,可检测因"窗口期"感染、免疫静默感染、病毒变异等原因而漏检的污染血液.因此,在许多发达国家和地区已普遍应用于献血者血液筛查,我国也有多个血液中心进行了血液NAT的评估性研究工作[1] 2010年11月本站开始应用核酸检测系统对常规血清学筛查合格的无偿献血者标本进行HBV、HCV和HIV的NAT检测,现报告如下.     

核酸检测技术在献血者血液筛查中的初步应用

目的将核酸检测(NAT)系统应用于献血者血液筛查过程。方法采用Roche Cobas S201系统对血站常规EIA检测合格献血者的65 497份标本进行HIV、HCV和HBV3项联合筛查,并对NAT筛查阳性的标本做确证试验。结果65 497份EIA检测合格标本中,NAT共检出阳性59例,阳性检出率为0.9‰;NAT筛查阳性标本经另1种核酸检测系统确证阳性率为65.38%(17/26)。结论NAT系统应用于献血者血液筛查有助于提高血液及输血安全。     

沧州地区合格献血者HBV、HCV和HIV核酸检测结果

输血后病毒感染的主要原因有：病毒感染者＂窗口期＂献血;病毒变异;免疫静默感染;以及人工操作错误,其中病毒感染者＂窗口期＂献血是主要原因。目前我国大部分血站采用2个不同厂家ELISA试剂筛查献血者传染病指标。由于该检测方法的局限性,使乙肝、丙肝、艾滋＂窗口期＂造成临床输血传播疾病风险依旧存在。核酸检测（NAT）是对病原体核酸直接检测,病毒感染数天即能检出极微量的病毒核酸,不仅能大大缩短＂窗口期＂且能避免免疫静默感染发生。     

2种检测模式及不同献血次数对献血者血液HIV项目检测后残余风险的影响

目的评估本地区无偿献血者经血液筛查后血液中HIV残余风险,分析其与检测策略和献血次数的关系。方法收集本站2014—2017年使用2种不同检测策略[ELISA单一试剂检测(丽珠,万泰,新创试剂盒)和ELISA双试剂检测(丽珠+万泰,万泰+新创试剂盒)]的实验室HIV抗原/抗体反应性结果及疾病预防控制中心(CDC)回报的确证结果,采用残余风险数学模型估算残余风险,分析其与检测策略和献血次数的关系。结果重复献血者的HIV流行率为0.024 3%(33/135 723),残余风险为1/699 212,初次献血者的HIV流行率为0.059 8%(108/180 587),残余风险为1/213 648,重复献血者的残余风险和流行率均低于初次献血者(P<0.05);单一试剂检测HIV的流行率为0.052 4%,残余风险为1/145 915,双试剂检测HIV的流行率为0.035 7%,残余风险为1/701 744,单一试剂的残余风险高于双试剂的残余风险(P<0.05)。结论进行HIV抗原/抗体不同ELISA试剂的残余风险估算可帮助实验室对不同的检测策略进行评价,筛选出适合本实验室的检测策略,提高检出率,同时为招募一些固定、低危的献血者提供依据,有利于保障血液安全。     

云南德宏州无偿献血者HIV HCV感染情况调查

云南省德宏州地处云南滇西边境,人口118万,少数民族占53.64%,与缅甸接壤,国境线长达503.8公里.特殊的地理条件使德宏吸毒贩毒形势严峻,是我国HIV高度流行的地区,同时又是我国HIV预防项目的重点地区.调查本州无偿献血人群HIV和HCV感染情况对进一步加强采供血工作,保证血液安全具有重要意义.为此我们对2006-2008年本州无偿献血者抗-HIV和抗-HCV检测情况进行调查分析,现报告如下.     

四川地区血站分离血浆HIV/HBV/HCV残余风险研究

目的评估四川地区血站分离血浆中HIV、HBV、HCV的残余风险。方法收集2013年1-12月四川9家血站HBs Ag、抗-HCV、抗-HIV筛查阳性样本,对筛查HBs Ag阳性标本采用中和试验确证,对筛查抗-HCV及抗-HIV阳性标本采用免疫印迹法确证。采用改良的发病率-窗口期模型计算HBV,HCV,HIV残余风险。结果四川地区血站分离血浆中HBV,HCV和HIV的残余风险分别为1/1 355,1/35 587和1/92 593。结论血站分离血浆仍存在一定残余风险,增加核酸检测等新型检测技术可进一步降低其残余风险。     

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。     

澳门地区献血者病毒核酸筛检的分析及跟进调查

目的了解澳门地区献血者标本中,HIV、HBV和HCV血清学检测为阴性,核酸检测为阳性的发生频率。方法采用回顾性调查方法,对2007年5月—2010年6月已经过病毒血清学检测(包括HBsAg、抗-HCV及抗-HIVⅠ/Ⅱ)、排除了病毒血清学筛查阳性后余下的40149个阴性献血者标本,做HIV、HBV及HCV的核酸筛检。结果共筛检测29个标本呈阳性反应,将它们做病毒区分检测:没有发现HIV及HCV阳性个案,确认为HBV阳性24例,其余5例为未能区分病毒种类。进一步追踪调查:在初筛确定为HBV核酸阳性的24例中,13例追踪标本HBV核酸检测呈阴性,2例(2例2次,1例3次)持续呈阳性。结论 1)澳门地区属乙型肝炎的高发区,实施病毒NAT筛检提高了血液的安全水平;2)追踪隐性乙型肝炎患者的HBV DNA变化和血清学检测,除了是关注献血者的健康状况外,亦为制定献血者筛除政策提供依据。     

结合核酸检测探讨ALT与HBV、HCV的相关性

目的探讨无偿献血者丙氨酸氨基转移酶(ALT)与HBV、HCV的相关性。方法分析本站2011年6月～9月共15 879份血液标本ALT、HBsAg、抗-HCV的检测结果,并对527例ALT≥60U/L的肝炎标志物阴性献血者标本进行NAT检测及相关性分析。结果 15 879份血液标本中,ALT异常者1 233份,其中合并HBsAg阳性或抗-HCV阳性者共28份,占全部ALT异常者的2.27%;肝炎标志物阳性组与肝炎标志物阴性组ALT异常率的差异均无统计学意义(P>0.05);527例ALT≥60 U/L的肝炎标志物阴性献血者NAT检测均为阴性。结论 ALT作为1项非特异性指标,与HBV和HCV检测结果的关联性无统计学意义。     

环介导的等温扩增技术对HBV、HCV和HIV核酸检测的研究

目的建立环介导的等温扩增技术(LAMP)对HBV、HCV和H IV核酸检测的方法并对检测灵敏度和特异性作初步考核。方法通过引物设计和筛选、内质控的设计与时间分辨浊度检测方法的运用,建立LAMP HBVDNA、HCV RNA和H IV RNA扩增体系;用连续稀释的阳性样本和不含任何病毒核酸的正常人血浆考核所建立的LAMP扩增体系的灵敏度和特异性。结果建立了LAMP HBV DNA扩增体系及HCV/H IV RNA双联检体系,该体系对5 CP/m l的HBV DNA样本的检出率为51.85%,对100 CP/m l的HCV RNA阳性样本的检出率为61.90%,对100 CP/m l的H IV RNA阳性样本的检出率为45%。对考核样本检测的相对灵敏度和特异性均为100%。结论LAMP HBV、HCV和H IV检测灵敏度较高,在进一步优化以后能用于HBV、HCV和H IV的核酸检测。     

Prevalence,incidence, and residual risk of human immunodeficiency virus and hepatitis C virus infections among United States blood donors since the introduction of nucleic acid testing

BACKGROUND: Nucleic acid testing (NAT) for human immunodeficiency virus (HIV) and hepatitis C virus (HCV) was introduced for blood donation screening in the United States in 1999. This study analyzes temporal trends of these two infections since NAT introduction. STUDY DESIGN AND METHODS: Donation data from 1999 to 2008 were analyzed; each donation was tested for antibodies and viral RNA for HIV and HCV. Incidence for first‐time (FT) donors was derived by multiplying that among repeat (RP) donors by the ratio of NAT yield rates between FT and RP donors. Incidence for all donors was the weighted mean based on percentage of FT and RP donors. Residual risk (RR) was determined using the window‐period model. RESULTS: During the 10‐year period approximately 66 million donations were screened with 32 HIV (1:2 million) and 244 HCV (1:270,000) NAT yield donations identified. HCV prevalence among FT donors decreased by 53% for 2008 compared to 1999. HIV and HCV incidence among RP donors increased in 2007 through 2008 compared to 2005 through 2006. During 2007 through 2008, HIV incidence was 3.1 per 10⁵ person‐years (py), with an RR estimate of 0.68 per 10⁶ (1:1,467,000) donations; HCV incidence was 5.1 per 10⁵ py, with an RR estimate of 0.87 per 10⁶ (1:1,149,000). The increase in HIV incidence was primarily among 16‐ to 19‐year‐old, male African American donors and that in HCV was primarily among Caucasian donors of 50 or more years. Donors from the Southern United States had higher incidence rates. CONCLUSION: HCV prevalence decreased significantly since NAT introduction. The increase in HIV and HCV incidence in 2007 through 2008 warrants continued monitoring and investigation.     

丙型肝炎病毒感染的献血者12年追踪观察

目的动态追踪观察HCV感染的献血者健康状况、疾病进程和转归。方法选择刚发生HCV感染的献血者,12年中定期抽血检测,观察其HCV RNA、抗-HCV和ALT的动态变化,并按HCV RNA出现的阳性频率和转阴趋势分为两组(第1组阳性率低且趋转阴,第2组则相反),调查这些感染者的健康状况,并进行肝脾B超检查和肝组织病理检查。结果①第1组中的7名受试者血清(血浆)HCV-RNA呈间隙性阳性,阳性率为41.6%(47/113),其血清ALT异常率为14.4%(16/111),其中4名作肝活检后均诊断为轻度慢性肝炎,病理改变程度是:G1、S0和G1、S1各2例;②第2组中的19名受试者血清(血浆)HCV RNA呈持续性阳性,阳性率为88.2%(276/313),其血清ALT异常率为50.3%(159/316),其中6名肝活检也诊断为轻度慢性肝炎,但病理改变明显,G2、S2有2例,G2、S1有4例;③26名受试者一般健康状况尚佳,除部分肝脾B超呈现实质回声增强外,无其他特殊情况发现。结论26名HCV感染的献血者12年间均已发展为轻度慢性丙型肝炎;其中7名受试者病情在恢复,19名受试者病情可能有发展。     

Impact of individual-donation nucleic acid testing on risk of human immunodeficiency virus, hepatitis B virus, and hepatitis C virus transmission by blood transfusion in South Africa

BACKGROUND: In 2005, the South African National Blood Service introduced individual-donation (ID) nucleic acid test (NAT) screening for human immunodeficiency virus (HIV) RNA, hepatitis C virus (HCV) RNA, and hepatitis B virus (HBV) DNA. At the same time the use of ethnic origin to prioritize the transfusion of blood according to a hierarchy of residual risk was discontinued.
STUDY DESIGN AND METHODS: ID-NAT (Ultrio on Procleix Tigris, Chiron) and serology (PRISM, Abbott) repeat test and confirmation testing algorithms were designed to enable differentiation between false-positive and true-NAT and -serology yields. After 1 year, the NAT and serology yield rates in first-time, lapsed, and repeat donors were analyzed and used to estimate the residual risk of HIV, HBV, and HCV infections by blood transfusion.
RESULTS: The HIV, HBV, and HCV ID-NAT window phase yield rates in 732,250 blood donations were 1:45,765, 1:11,810, and 1:732,200, respectively. Seven of 16 HIV window phase donations with viral loads above 16,000 copies/mL were HIV p24 antigen enzyme-linked immunosorbent assay positive. PRISM detected anti-HIV and hepatitis B surface antigen (HBsAg) in 89.4 and 73.9% of early infections in repeat donors. The Procleix assay detected viremia in 99.7 and 95.5% of anti-HIV– and HBsAg-positive first-time donors. In these donors, the occult HBV DNA carrier rate was 1:5200. The residual transmission risk of ID-NAT HIV, HBV, and HCV window phase donations was estimated at 1:479,000, 1:61,500, and 1:21,000,000 respectively.
CONCLUSION: One-year ID-NAT screening of 732,250 donations interdicted 16 HIV, 20 HBV, and 1 HCV window phase donations and 42 anti-hepatitis B core antigen–reactive infections during an early recovery or a later stage of occult HBV infection.     

Antibody to hepatitis C virus in blood donors found positive for other agents. II. Anti-HIV-1

Summary. Stored serum samples from 24 blood donors confirmed positive for anti-HIV-1 were tested for antibody to hepatitis C virus (HCV). Those repeatedly reactive using the anti-HCV ELISA screening test were retested by the HCV recombinant immunoblot (RIBA). Risk-factors for the contraction of HIV infection that had been elicited at formal counselling sessions were evaluated in relation to HCV/HIV modes of infection. The only two donors confirmed to be anti-HCV positive both admitted to intravenous drug use.     

The cost‐effectiveness of introducing nucleic acid testing to test for hepatitis B,hepatitis C,and human immunodeficiency virus among blood donors in Sweden

BACKGROUND: The purpose of this study was to estimate the cost‐effectiveness of using individual‐donor nucleic acid testing (ID‐NAT) in addition to serologic tests compared with the sole use of serologic tests for the identification of hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) among blood donors in Sweden. STUDY DESIGN AND METHODS: The two strategies analyzed were serologic tests and ID‐NAT plus serologic tests. A health‐economic model was used to estimate the lifetime costs and effects. The effects were measured as infections avoided and quality‐adjusted life‐years (QALYs) gained. A societal perspective was used. RESULTS: The largest number of viral transmissions occurred with serologic testing only. However, the risks for viral transmissions were very low with both strategies. The total cost was mainly influenced by the cost of the test carried out. The cost of using ID‐NAT plus serologic tests compared to serologic tests alone was estimated at Swedish Krona (SEK) 101 million (USD 12.7 million) per avoided viral transmission. The cost per QALY gained was SEK 22 million (USD 2.7 million). CONCLUSION: Using ID‐NAT for testing against HBV, HCV, and HIV among blood donors leads to cost‐effectiveness ratios that are far beyond what is usually considered cost‐effective. The main reason for this is that with current methods, the risks for virus transmission are very low in Sweden.