江苏地区无偿献血者HCV基因型的分布

目的探讨江苏地区献血者感染的HCV基因型及亚型。方法收集2013年来自江苏省血液中心ELISA双试剂检测抗-HCV阳性献血者血清标本,使用HCV核酸定量检测试剂盒检测HCV RNA载量;同时使用RNA提取试剂提取HCV RNA,反转录PCR法扩增HCV Core区基因片段,并对扩增产物进行测序,利用进化树分析基因型和亚型。结果 2013年抗-HCV阳性标本139例(0.20%),其中64例抗-HCV阳性血清标本经荧光定量检测,RNA阴性30份,阳性34份。经巢式PCR扩增能够分型的标本24份,包括1a(71.7%)、1b(7.5%)、2a(7.5%)和3b(1.9%)3种基因型和4种亚型。年龄偏大(35岁)以及男性的HCV RNA阳性标本的病毒滴度与抗-HCV水平(S/CO值)都高于RNA阴性标本,并且存在性别差异,但年龄间无差异。结论江苏无偿献血者感染的HCV基因型以1型为主,其中1b为优势基因亚型。病毒血症献血者的HCV抗体水平显著升高,HCV RNA在自然感染进程中的变化有待通过进一步随访进行研究。     

广州地区首次参加无偿献血人群HCV感染的流行病学调查

目的了解广州地区首次参加无偿献血人群HCV的感染状况。方法抗-HCV检测采用酶联免疫吸附实验(使用2种不同厂家的试剂盒),HCV RNA检测采用荧光定量PCR法,ALT检测采用速率法。结果2004—2007年在广州血液中心首次参加无偿献血的559890名献血者中,1617名经双试剂检测抗-HCV阳性,阳性率为0.289%。随机选取435名双试剂检测抗-HCV阳性血液标本进行RNA检测,266名阳性,占61.15%,男女(P<0.01)HCV RNA阳性率有统计学意义(P<0.01)。266名抗-HCV检测阳性献血者中,ALT升高(>40U)的为18名(其中男性17名),不合率为6.77%,男女ALT的不合格比有统计学意义(P<0.01)。结论随着无偿献血的开展,献血人群中HCV的阳性率不断下降;不同性别的个体清除HCV的能力不同,女性高于男性。     

深圳献血者中HCV感染自然清除与病毒血症特征分析

目的测定我国深圳地区献血者中的丙型肝炎病毒(hepatitis C virus,HCV)感染者自然清除和病毒血症,即自然康复与慢性感染者比率及其人群特征,为丙型肝炎防治研究提供数据。方法对深圳献血者中抗-HCV初筛阳性的血清标本采用2种EIA方法进行抗体再测定,以定量PCR方法测定病毒载量,并采用巢式PCR对核酸进行确认,进而将标本分为3种HCV感染状态,即病毒自然清除(RNA-/Ab+)、病毒血症(RNA+/Ab+)和假阳性(RNA-/Ab-),通过统计学方法分析3种感染状态献血者在临床信息(性别、年龄)、ALT、抗-HCV的差异。HCV RNA阳性标本通过分析5'-NCR序列进行基因分型。结果 152份初筛抗-HCV阳性的标本中,病毒自然清除标本45份、病毒血症51份、假阳性56份。50份HCV定量PCR阳性标本病毒载量范围从[(12.6～2.43)×106]IU/ml(中位值2.54×104IU/ml)。36份进行基因分型的标本包括47.2%基因1型、5.6%基因2型、19.4%基因3型和27.8%基因6型。慢性感染组标本的年龄及抗-HCV水平(S/CO值)显著高于病毒自然清除和假阳性组(χ2=7.812,P0.05;χ2=90.865,P0.01)。结论深圳地区献血者HCV感染病毒自然清除率约为46.9%。HCV基因型中1型为主要,6型也占有较高比例。年龄小、女性献血者更易于自然清除病毒。病毒血症即慢性感染者HCV抗体水平显著升高,其在HCV自然康复过程中的作用还有待进一步研究。     

广州市无偿献血者中抗-HCV阳性人群的比较分析

目的了解广州市不同性别、年龄以及不同职业的无偿献血者抗-HCV阳性率情况。方法对从2000年1月～2010年12月的2 520 179(人)份广州市无偿献血者的血液标本,采用ELISA做抗-HCV双试剂检测并对检测结果做统计学分析。结果广州地区无偿献血者抗-HCV阳性率为0.42%(10 570/2 520 179),且在不同性别、年龄、职业之间差异具有统计学意义(P0.05)。女性抗-HCV阳性率(0.29%)低于男性(0.52%);35岁[18～25岁(0.40%);25～35岁(0.36%)]较≥35岁[35～45岁(0.54)%,45～55岁(0.52%)]的献血者的抗-HCV阳性率低;工人(0.56%)和无职业者(0.76%)的抗-HCV阳性率比较高,医务人员血样中未检出抗-HCV阳性。结论广州市无偿献血人群中抗-HCV阳性率较低,符合献血条件的≤35岁的人群积极献血将有利于保障临床输血安全;宜加强对工人和无职业者献血安全和输血相关病毒知识的宣传和教育。     

2012～2014年无偿献血者抗-HCV筛查结果分析

目的了解盐城地区献血者丙型肝炎病毒(HCV)感染情况。方法采用酶联免疫吸附法(ELISA)检测献血者抗-HCV,用荧光定量聚合酶链反应检测HCV RNA。结果盐城地区献血者抗-HCV阳性率为0.07%(109/163 782);109例抗-HCV阳性者中,80例为单试剂阳性,29例为双试剂阳性;初次献血者抗-HCV阳性80例,重复献血者抗-HCV阳性29例;符合核酸检测要求的72例单试剂抗-HCV阳性者均为HCV RNA阴性。结论盐城地区献血者抗-HCV阳性率低于普通人群,近三年献血者抗-HCV阳性率没有变化,抗-HCV单试剂阳性者核酸检测均为阴性。     

深圳市无偿献血者抗-HCV阳性结果分析

目的了解深圳市无偿献血人群中丙型肝炎病毒(HCV)的感染情况,探讨HCV酶联免疫吸附试验(ELISA)检测与HCV RNA及丙氨酸氨基转移酶(ALT)结果的相关性,为献血者的招募提供依据,降低输血风险。方法对从2014年1月至2015年10月的深圳市无偿献血标本进行抗-HCV ELISA、ALT及HCV RNA检测,对相关数据进行分析,并对抗-HCV阳性数据从不同年龄、性别、不同职业、不同献血次数等方面进行统计分析。结果 249 585份标本中共检测出抗-HCV阳性480例,阳性检出率为0.19%,其中ALT异常2例,HCV RNA阳性104例。女性抗-HCV阳性构成比低于男性。不同职业献血者的抗-HCV阳性率不同,医务人员、学生、军人等检出率相对较低。重复献血者感染率明显低于初次献血者,差异有统计学意义(P0.01)。结论深圳市无偿献血人群HCV感染率较低,随着核酸检测的开展,ALT检测在无偿献血筛查中的作用值得进一步讨论。建议对工人及其他职业人群加强健康宣传教育,针对低危人群建立相对固定的无偿献血队伍,保障临床用血安全。     

茂名地区无偿献血者72例HCV感染特征及检测效果评价分析

目的测定茂名地区部分丙型肝炎病毒感染献血者中不同感染状态比率及其分子特征,分析丙型肝炎病毒感染基因型分布流行特征及趋势,评价ELISA法检测血液HCV的效果。方法对本地区献血者中抗-HCV检测呈反应性的血液标本采用定量PCR方法检测血浆中病毒载量,同时采用巢式PCR技术扩增核酸片段,进而将血液标本分为病毒自然清除(RNA-/抗-+)与病毒血症(RNA+/Ab抗-+)2种状态,并分析2种状态在献血者性别方面的差异。HCV RNA阳性标本通过分析5’-NCR序列进行基因分型。比较ELISA与NAT检测方法的符合性。结果72份抗-HCV呈反应性的标本中,病毒自然清除50份(69.4%),病毒血症22份(30.6%)。22份PCR定量阳性标本中基因分型阳性17份,其中基因1型占17.6%(3/17),基因2型占11.8%(2/17),基因3型17.6%(3/17),基因6型47.1%(8/17),无法准确定型5.8%(1/17)。试剂A检测呈反应性标本中核酸确认阳性标本占32.3%;试剂B为54.5%。女性病毒血症的比例(2/19)比男性(20/53)低27.2%(χ2=9.637,P0.05)。结论茂名地区献血者HCV感染病毒自然清除率约为69.4%,HCV基因型中6型为主要基因型,女性献血者更容易清除病毒。现行2遍ELISA检测策略安全有效,但同时存在特异性不高的弊端,实施ELISA加NAT检测的策略,更能保证血液的安全和降低检测假阳性。     

实时荧光定量PCR筛查献血者HCVRNA的初步研究

目的比较核酸扩增技术(NAT)和血清学酶联免疫试验(ELISA)对献血者丙型肝炎病毒(HCV)筛查的符合率。方法用实时荧光定量PCR方法对ELISA初、复检抗-HCV均为阳性的献血者血浆和抗-HCV阳性的丙型肝炎患者血浆作HCV RNA检测。结果①36例初、复检抗-HCV均为阳性的献血者血浆为HCV RNA13例阳性,答合率36.11%,31例抗-HCV阳性的丙型肝炎患者血浆的HCV RNA检测为24例阳性,符合率77.42%,两者符合率有显著性差异(χ2=11·49,P<0·005);②前述37例HCV RNA阳性血浆分别与23份初、复检抗-HCV和HCV RNA均为阴性的血浆混合后HCV RNA检测仍为阳性。且其中1例阳性血浆用上述阴性血浆100倍稀释后检测仍为阳性;③70,000例沈阳地区初、复检抗-HCV阴性献血者血浆样本NAT混合法筛查均为阴性。结论NAT可作为血清学ELISA法的补充用于对献血者血液HCV的常规筛查。     

血浆miR-122在抗-HCV阳性献血者中表达的动态分析

目的动态分析抗-HCV阳性献血者血浆miR-122的表达特点及与HCV感染诊断指标的关联。方法对133名抗-HCV阳性献血者、24名健康献血者及15名确证抗-HCV阳性追踪献血者血液标本抽提血浆miRNA,以实时荧光定量PCR法检测血浆miR-122,分析血浆miR-122相对表达量与HCV RNA、抗-HCV临界值指数(COI)等指标关联性。结果 ROC曲线分析:miR-122在抗-HCV+/HCV RNA+组和抗-HCV+/HCV RNA-组中诊断丙型肝炎的AUC(ROC曲线下面积)为0.830(95%CI:0.755-0.889),区分抗-HCV+/HCV RNA+组和抗-HCV+/HCV RNA-组的敏感性和特异性分别为74.7%和82.4%。追踪15例抗-HCV+/HCV RNA+献血者血浆miR-122表达明显降低(P0.05)。结论血浆miR-122与HCV感染密切相关,确证HCV RNA感染献血者血浆miR-122随时间迁移表达降低。     

深圳地区抗-HCV阴性/NAT初筛阳性献血者的HCV窗口感染期确认

目的了解深圳地区血液筛查中抗-HCV阴性/NAT初筛阳性献血者的检出情况及其HCV窗口感染期的确认。方法 2008-2014年本中心采用酶免(ELISA)及病毒核酸检测方法(NAT)筛查492 325份无偿献血者样品,获得6例抗-HCV阴性/NAT初筛阳性献血者,使用荧光定量检测方法(QPCR)和巢式PCR(Nested-PCR)确认其HCV RNA是否存在并随访跟踪复查,以确定6例献血者核酸检测结果假阳性或HCV窗口感染期的性质。结果本中心在这7年期间,492 325份献血者的抗-HCV阴性/NAT初筛阳性检出率为1/82 054(6/492 325);6例抗-HCV阴性/NAT初筛阳性献血者,确认1/6例(16.7%)处于HCV窗口感染期,1/6例(16.7%)判定为核酸检测采血管血样污染HCV造成假阳性结果,4/6例(66.7%)判定为核酸仪器检测过程中出现假阳性导致NAT初筛阳性。故本中心目前献血者HCV窗口感染期检出率为1:492 325。结论目前核酸检测存在假阳性情况,需建立额外核酸扩增方法及追踪随访才能确定样品性质,确保病毒感染窗口期结果的准确性。     

化学发光微粒子免疫测定技术对低风险人群HCV感染确证的应用价值

目的应用化学发光微粒子免疫测定(CMIA)技术检测一种酶联免疫吸附试验(ELISA1)反应性样本,通过HCV核酸和蛋白检测参考标准分析CMIA在献血者HCV感染确证中的应用价值。方法 102例ELISA1反应性献血者血液样本补充核酸3项联检、另一种酶联免疫吸附试验(ELISA2)、丙型肝炎病毒抗体补充试验(Western Blot法)和CMIA试验。结果 102例抗-HCV阳性献血者中,32例(31.37%,32/102)HCV RNA反应性样本,50例(49.02%,50/102)ELISA2及Western Blot均为反应性。以HCV核酸检测结果为参考标准,CMIA与之低度相关(Spearman相关系数rs=0.395,P0.01),Kappa检验两者一致性弱(Kappa=0.270,P0.01)。以ELISA2及Western Blot蛋白检测结果为参考标准,CMIA与之结果高度相关(Spearman相关系数rs=0.713,P0.01),Kappa检验两者高度一致性(Kappa=0.674,P0.01)。结论 CMIA作为HCV感染后蛋白标记物的检测方法,对低风险人群HCV病毒感染的确证有较大的应用价值。     

Hepatitis C in blood transfusion recipients identified at the Oxford Blood Centre in the national HCV look-back programme

After the introduction in September 1991 of donor screening for hepatitis C, 95 potentially infectious blood donors who had given blood before this date were identified at the Oxford blood centre. Three hundred and ninety-nine blood components issued previously from these donors were identified in the course of the national HCV look-back programme. Of 399 questionnaires sent to hospital blood banks 392 were returned, identifying 290 recipients of whom 177 (61%) had died, and 113 (39%) were still alive 4–13 years after transfusion. One hundred and four recipients were traced and tested. Forty-nine recipients were not HCV infected. Forty-four of 58 (76%) recipients who received blood from donors found to be HCV RNA positive after September 1991 gave positive test results for HCV RNA. Eleven of 58 showed only antibody (anti-HCV), and 3/58 who had apparently received infectious blood showed no sign of past infection. The 11 who showed anti-HCV only, together with the three who showed no sign of past infection despite strong evidence of receiving HCV RNA-positive blood, had a mean age at transfusion of 27 years, compared with mean age at transfusion of 46 years in the 44 recipients with persistent HCV infection. Virus genotyping in 33/44 HCV RNA-positive recipients revealed five different genotypes. These did not seem to influence the outcome. Virus genotypes in 31 donor–recipient pairs showed complete concordance. Liver biopsies in 23/44 RNA-positive recipients showed minimal inflammation in four, mild in eight and moderate in 11. Liver fibrosis, Ishak grades 1–3, was present in 16/23 recipients. One other male recipient, not subjected to a liver biopsy, developed a hepatocellular carcinoma which caused his death at the age of 71, 8 years after transfusion.     

上海地区抗-HCV反应性献血者随访研究

目的对抗-HCV反应性献血者进行随访以分析反应性献血者的归队途径。方法随机对上海地区52名抗-HCV单试剂反应性献血者献血间隔6个月后随访,使用4种采供血机构常用的抗-HCV ELISA试剂检测,对于抗-HCV反应性标本使用重组免疫印迹试验确证,同时使用罗氏cobas Taqscreen MPX核酸检测试剂(NAT)单人份检测;3～6个月后进行第2次随访,开展相同实验。结果 2次随访研究发现,52名献血者中,39名(75%)献血者4种抗-HCV试剂检测结果均为阴性,13名(25%)献血者4种抗-HCV试剂至少有1种试剂检测结果为反应性,与原抗-HCV试剂检测结果相同,且S/CO数值保持基本一致,13名抗-HCV反应性献血者中有1名免疫印迹检测结果为阳性;52名献血者NAT结果均为阴性。结论献血者抗-HCV单试剂反应性,经过至少6个月间隔,使用包含原抗-HCV在内的2种试剂检测结果阴性,同时NAT检测阴性者可恢复献血权利。     

Characteristics of hepatitis C-positive blood donors in Victoria, Australia

Data from 3156 voluntary blood donors in Victoria, Australia, who were repeatedly reactive on anti-HCV screening test were analysed. It showed that between 1990 and 1997, 0.97% of routine blood donors were anti-HCV repeat reactive. Sixty-four (64) donors tested positive for HCV RNA. They were more likely to be male, had a mean age of 32.7 +/- 10.9 years and a mean serum ALT level of 70.9 +/- 46.5 i.u.L-1: the latter value being significantly different from donors with biological false reactivity (P < 0.0001). Among HCV antibody-positive donors there was a significantly increased rate of prior injecting drug use (34 vs. 1%), transfusion (25 vs. 10%) and tattooing (31 vs. 7%) than among donors with biological false reactivity (all P < 0.0001). The association of these risk factors is stronger with positivity for both immunoblot and HCV RNA than immunoblot alone. The data suggest that efforts should be made to increase the efficacy of donor questioning about prior drug use.     

Impact of screening blood donors for hepatitis C antibody on posttransfusion hepatitis: a prospective study with a second-generation anti-hepatitis C virus assay

BACKGROUND: Hepatitis C is the major cause of posttransfusion hepatitis. Blood components that are positive for antibody to hepatitis C virus (anti-HCV) can transmit posttransfusion hepatitis. STUDY DESIGN AND METHODS: To investigate the effect on posttransfusion hepatitis of screening blood donors with a second-generation test for anti-HCV, 249 transfusion recipients who underwent cardiovascular surgery were prospectively followed. Six recipients who were positive for anti-HCV before transfusion and 51 subjects with incomplete follow-up were excluded from this study. RESULTS: Eleven (13.8%) of 80 subjects who received unscreened blood had two successive serum alanine aminotransferase levels > 90 U per L. Seven (8.8% of total) developed anti-HCV and HCV RNA and two (2.5% of total) developed IgM antibody to cytomegalovirus (IgM anti-CMV). By contrast, 3 (2.7%) of the 112 subjects who received anti-HCV-screened blood had two successive serum alanine aminotransferase levels > 90 U per L. None of these three developed anti-HCV and HCV RNA, but two (1.8% of total) showed the development of IgM anti-CMV. The study shows that screening for anti- HCV in blood donors with a second-generation test almost abrogated posttransfusion viral hepatitis C. CONCLUSION: After anti-HCV screening, other body fluid-transmitted viruses such as CMV may become important in posttransfusion hepatitis.     

Outcome of post-transfusion hepatitis C: disease severity in blood- component recipients and their implicated donors

The UK 'Look-back Program' identifies recipients of blood products from hepatitis C antibody (anti-HCV) positive donors. Of 60 such recipients tested by the Newcastle Transfusion Service, 28(46.7%) were anti-HCV- negative, 25(41.7%) were anti-HCV-positive, and seven (11.6%) had equivocal serology. We studied 29 anti-HCV-positive/indeterminate recipients and eight of their implicated donors, using serial liver function tests (LFTs), liver histology when clinically indicated, HCV RNA and serotyping. Presumed resolved hepatitis C, with persistently normal LFTs and negative HCV RNA, was found in 28%, of whom 63% had indeterminate anti-HCV by RIBA (1 band of 4 detected on third- generation recombinant immunoblot assay). Resolved hepatitis C was significantly more common in women (p < 0.05) and tended to be associated with younger age at transfusion. There was complete concordance in serotype between donor-recipient pairs. There was no correlation in disease severity between recipients and their implicated donors, nor between recipients from the same donor. A history of alcohol consumption above recommended 'safe' limits (median 30 units) was associated with more severe histological disease (p < 0.01). Host factors, including gender and alcohol consumption, may be important in determining the wide variability in outcome of post-transfusion hepatitis C.      

Seroprevalence of hepatitis C virus (HCV) infection among blood donors: a hospital-based study.

Insofar as chronic hepatitis C virus (HCV) infection in many individuals is asymptomatic, and as the prevalence of antibodies to hepatitis C virus (anti-HCV) among blood donors in Lebanon is scarce, this study addressed the prevalence of anti-HCV in 5,115 blood donors. Data obtained were compared to other world regions. Of the blood donors screened, 57 were initially tested positive or doubtful for anti-HCV Ab. Subsequent testing by two-third generation enzyme immunoassays confirmed that, of the 57 initially tested positive/doubtful, only 18 were positive for anti-HCV giving a prevalence rate of 0.4%. While there was no difference in HCV prevalence with respect to age or gender, a higher rate was seen in non-Lebanese compared to Lebanese subjects (3.4% vs 0.3%, P < 0.001). These results demonstrate a low prevalence of HCV infection among Lebanese blood donors, which was comparable to those established for western countries.     

Hepatitis C virus among blood donors: follow-up study

BACKGROUND: The exact significance of antibodies to hepatitis C virus (HCV) in blood donors remains unknown. Confirmatory tests of anti-HCV- reactive serum and HCV RNA by polymerase chain reaction (PCR) are used to refute a large proportion of false-positive results. STUDY DESIGN AND METHODS: Ninety-two blood donors who were anti-HCV reactive in a first-generation enzyme-linked immunosorbent assay (ELISA) were reevaluated 10 months later with a second-generation ELISA (ELISA-2) as well as with second-generation recombinant immunoblot assay (RIBA-2) and by PCR. RESULTS: Twenty-five (43.9%) of the 57 ELISA-2-positive donors were confirmed as positive by RIBA-2; of these, 84 percent were HCV RNA positive in PCR. Of the 57 who were still anti-HCV positive, 46 were followed up and tested again in the same manner 2 years after the first screening. At that time, the pattern was little changed: 94 percent of RIBA-2- and PCR-positive donors remained positive. Of RIBA-2- and PCR-positive blood donors, 62 percent had abnormal alanine aminotransferase levels in at least one of the three evaluations. Among the anti-HCV-positive donors confirmed by RIBA-2, 60 percent, versus 12.6 percent in the control group, had a significantly (p < 0.001) more frequent risk factor for HCV infection, due to parenteral exposure to blood. CONCLUSION: These data confirm a good correlation between RIBA-2 reactivity and the detection of HCV RNA in a population of anti-HCV- positive blood donors.