血液病患者输血与庚型肝炎病毒感染的研究

目的：了解血液病患者输血后的庚型肝炎病毒（ＨＧＶ）感染情况。方法：对６３例血液病患者采用ＲＴＰＣＲ方法检测ＨＧＶＲＮＡ、丙型肝炎病毒（ＨＣＶ）ＲＮＡ，应用ＥＬＩＳＡ方法检测抗ＨＧＶ、ＨＢｓＡｇ。结果：ＨＧＶＲＮＡ阳性率为７．９％，抗ＨＧＶ阳性率６．３％；ＨＣＶＲＮＡ阳性率为４６．０％。ＨＧＶ感染通常伴有ＨＣＶ感染及丙氨酸转氨酶升高。ＨＧＶ感染与输血量有关，与血液病种类无关。结论：血液病输血可引起ＨＣＶ、ＨＧＶ的传播     

血透患者检测抗HCV IgM的临床意义

为探讨丙型肝炎ＩｇＭ抗体测定在血液透析患者中的临床意义,采秀间接ＥＬＩＳＡ法检测抗ＨＣＶ ＩｇＭ。同时采用ＥＬＩＳＡ测抗ＨＣＶ ＩｇＧ,ＲＴ－ＰＣＲ法测ＨＣＶ ＲＮＡ,并进行比较。结果示６２例血液透析病人中,抗ＨＣＶ ＩｇＭ阳性２７例（４３．６％）,抗ＨＣＶ ＩｇＧ阳性２９例（４６．８％）,ＨＣＶ ＲＮＡ阳性３４例（５４．８％）,任一项阳性３７例（５９．７％）;抗ＨＣＶ ＩｇＭ与ＨＣＶ ＲＮＡ检测     

第三代酶免疫试验检测丙型肝炎病毒抗体的意义

本文应用Ａｂｂｏｔｔ公司第三代酶免疫试验（ＥＩＡ－３），与Ａｂｂｏｔｔ公司和国产第二代酶免疫试验（ＥＩＡ－２），对比检测了９６例肝炎病人和献血员标本的抗－ＨＣＶ，发现Ａｂｂｏｔｔ公司和国产ＥＩＡ－２的相对漏检率分别为１７．３％和２０．７％，而献血员中有漏检１０．８％。对部分漏检标本应用多聚酶链试验（ＰＣＲ）检测了ＨＣＶ－ＲＮＡ，发现与ＨＣＶ感染有关。这些是造成某些临床误诊及输血后肝炎的重要原因之一     

复合PCR检测乙型肝炎和丙型肝炎病毒

设计了ＨＢＶ和ＨＣＶ各一对引物，建立了复合ＰＣＲ，并对临床标本进行检测，５例ＨＢｓＡｇ和ＨＣＶ抗体均阳性者，其ＨＢＶ－ＤＮＡ和ＨＣＶ－ＲＮＡ均阳性；２０例单ＨＢｓＡｇ阳性标本，１１例单ＨＢＶ－ＤＮＡ阳性，５例ＨＢＶ－ＤＮＡ和ＨＣＶ－ＲＮＡ均阳性；２０份单ＨＣＶ抗体阳性者，ＨＣＶ－ＲＮＡ均阳性，３例合并ＨＢＶ－ＤＮＡ阳性；１０例慢性ＮＡＮＢ抗ＨＣＶ阴性肝炎标本８份ＨＣＶ－ＲＮＡ，１例ＨＢＶ－ＤＮＡ和     

输血及血制品者丙型肝炎病毒感染调查

对深圳九龙海关职员有输血或血制品史者６６铭及对照组１４７名调查其ＨＣＶ感染情况。结果：输血组抗ＨＣＶ组性率为９．１％，明显高于抽样组的０．７％。输血组中以血浆者抗－ＨＣＶ检出最高，传播风险较大。丙种球蛋白及补血康输入者抗－ＨＣＶ阴性。抗－ＨＣＶ检出阳性者中ＡＬＴ升高者ＨＣＶＲＮＡ均阳性，提示ＨＣＶＲＮＡ水平与活动性肝损害相关。     

逆转录聚合酶链反应检测血清中丙型肝炎病毒RNA

本文用本所自己合成的引物，对３０份怀疑ＨＣＶ感染血清进行ＨＣＶＲＮＡＲＴ－ＰＣＲ检测。该３０份血清中，２２份抗－ＨＣＶ阳性，其中１８份ＨＣＶＲＮＡ为阳性，８份抗－ＨＣＶ阴性血清及怀疑阳性血清中，２份ＨＶＣＲＮＡ为阳性。结果提示，抗－ＨＣＶ抗体试剂盒用于检测急性ＨＣＶ感染有其不足之处，结合ＰＣＲ检测ＨＣＶＲＮＡ的方法可提高急性ＨＣＶ感染的检出率。     

干扰素治疗前丙肝患者血清及外周血单个核细胞中HCV—RNA的检测

本文采用ＲＴ－ＰＣＲ方法对２６例干扰素治疗前丙肝患者血清ＨＣＶ－ＲＮＡ进行检测，并对１４例血清ＨＣＶ－ＲＮＡ阴性的标本进行外周血单个核细胞中ＨＣＶ－ＲＮＡ进行检测，发现２６例血清标本中ＨＣＶ－ＲＮＡ阳性率为２６．９％，而１４例血清ＨＣＶ－ＲＮＡ阴性标本，其ＰＢＭＣ中ＨＣＶ－ＲＮＡ有３例阳性，阳性率为２１．１％。因此，ＰＢＭＣ中ＨＣＶ－ＲＮＡ是丙肝病毒血症的又一个诊断指标，是否可作为干扰素疗效判断的     

TthDNA聚合酶在丙型肝炎病毒RNA检测中的应用

用ＴｔｈＤＮＡ聚合酶行逆转录聚合酶链反应（ＲＴ－ＰＣＲ）扩增丙型肝炎病毒（ＨＶＣ）５’端非编码序列，对２６２例肝炎患者血清进行ＨＣＶＲＮＡ测定，结果ＨＣＶＲＮＡ阳性者５３例，阳性率２０．２％。凡阳性者用酶联免疫吸附法测其抗ＨＣＶＩｇＧ，其中３０例测到抗ＨＣＶ，阳性重叠率５６％（３０／５３）；ＨＣＶＲＮＡ阳性中的１５例同时用成髓细胞瘤病毒酶行传统逆转录。套式聚合酶链反应检测对照，１４例阳性，符合率达９３．３％（１４／１５）。ＴｔｈＤＮＡ聚合酶用于ＲＴ－ＰＣＲ检测ＨＣＶＲＮＡ５’端非编码序列片段具有敏感性高、特异性强、稳定性好的优点。     

逆转灵—套式聚合酶链反应检测丙型肝炎病毒RNA

采用逆转录－套式聚合酶链反应检测２８０例疑为ＨＣＶ感染的病人血清，并同时做抗－ＨＣＶ－ＩｇＧ检测。结果５１例ＨＣＶ－ＲＮＡ阳性，其中２６例抗ＨＣＶ－ＩｇＧ阳性；在２５４例抗ＨＣＶ－ＩｇＧ阴性血清中有２５例ＨＣＶ－ＲＮＡ阳性。     

肝组织HCV—NS3抗原与HCV感染血清不同模式的关系及临床意义

采用逆转录套式聚合酶链反应（ＲＴ－ｎｅｓｔｅｄ ＰＣＲ）和ＥＬＩＳＡ方法检测血清ＨＣＶ ＲＮＡ、抗－ＨＣＶ，采用鼠单克隆抗体ＰＡＰ方法检测肝组织内ＨＣＶ－ＮＳ３抗原。结果发现被检测的４５例丙型肝炎患者随着病情加重和病程延长，血清抗－ＨＣＶ和ＨＣＶ ＲＮＡ及肝组织ＨＣＶ－ＮＳ３抗原阳性率逐步增高；某些患者抗－ＨＣＶ和ＨＣＶ ＲＮＡ阳性结果不一致；血清ＨＣＶ ＲＮＡ阳性肝组织ＨＣＶ－ＮＳ３阳性率明显同     

冻干低pH静注免疫球蛋白传播丙型肝炎病毒危险性观察与探讨

通过对冻干低ｐＨ静注免疫球蛋白成品的检测和临床使用观察，发现对献血者进行抗－ＨＣＶ抗体筛选，可使成品抗－ＨＣＶ阳性率大幅度降低。由于筛选方法的限制和ＨＣＶ感染后空窗期的存在，合并后的原料血浆污染ＨＣＶ在所难免；冻干低ｐＨ静注免疫球蛋白成品无论其抗－ＨＣＶ是阴性还是阳性，ＰＣＲ检测ＨＣＶ－ＲＮＡ均为阴性的实验结果提示制备工艺过程可以去除ＨＣＶ。临床初步观察和回顾性调查中均未发现ＨＣＶ传播的迹象。     

Efficacy of HCV core antigen detection during the preseroconversion period

BACKGROUND: The purpose of this study was to compare the performances of HCV core antigen (HCV Ag) testing with HCV RNA detection during the preseroconversion period. STUDY DESIGN AND METHODS: Six HCV antibody (HCV Ab)-negative and HCV RNA-positive blood samples from 6 donors and 135 serial samples from 28 patients who had undergone hemodialysis, collected a mean of 90 days before the detection of HCV Ab, were tested by ELISA for the detection of HCV Ag and by PCR to quantify HCV RNA. RESULTS: Five of the six donors were positive for HCV Ag. The donor with a negative HCV Ag test had the lowest viral load. In the hemodialysis patients, the 43 first specimens of the series were HCV RNA negative. Of the 92 specimens that were HCV RNA positive, 81 (88%) were positive for HCV Ag. Among the 74 samples with more than 10(5) RNA copies, 71 (96%) were HCV Ag positive. Average time from first viremic bleed to first HCV Ag-positive bleed was estimated at 2.0 days and that to first HCV Ab-positive bleed at 50.8 days. CONCLUSION: HCV Ag testing permits the detection of an HCV infection about 1.5 months earlier than the HCV Ab screening tests and an average of only 2 days later than quantitative HCV RNA detection in individual specimens.     

抗-HCV结果分析及灰区范围设置的探讨

目的分析酶联免疫吸附试验(ELISA)筛查丙型肝炎病毒抗体(抗-HCV)结果,探讨实验室ELISA中灰区范围设置的必要性。方法回顾性分析34 942例患者抗-HCV筛查结果,比较抗-HCV与HCV-RNA及临床确诊丙型肝炎患者间的关系;以初筛S/CO值在0.4~2.0范围内的标本为灰区样本,进行不同厂家试剂复检及HCV-RNA检测,探讨设置抗-HCV检测灰区范围的必要性。结果抗-HCV筛查阳性率0.61%;31~50岁阳性率最高;男性高于女性,两者比较差异有统计学意义(P0.05)。S/CO≥10时抗-HCV与HCV-RNA检测结果符合程度高,S/CO≥3.8时抗-HCV与临床确诊丙型肝炎符合程度高。灰区样本的阳性率0.38%,双试剂双孔复检后阳性率0.20%和0.05%。结论抗-HCV筛查阳性率在不同地域、性别及年龄段存在差异;S/CO值越大,HCVRNA阳性率越高,与临床丙型肝炎的确诊符合程度越高,而抗-HCV筛查落在灰区范围的样本应复检并检测RNA,以减少实验室漏检或假阳性结果的产生。     

Laboratory diagnosis and molecular epidemiology of an outbreak of hepatitis C virus infection among recipients of human intravenous immunoglobulin in Spain

BACKGROUND: Passive transfer of antibody to hepatitis C virus (HCV) has been thought to occur after infusion of human intravenous immunoglobulin (IVIG), as anti-HCV and/or HCV RNA was commonly found in that product. Recently, however, HCV RNA was detected in the serum of recipients of IVIG. Establishment of a causal relationship between IVIG therapy and HCV infection in recipients was attempted. STUDY DESIGN AND METHODS: Anti-HCV and HCV RNA sequences were investigated in serum samples from 39 persons who received a human IVIG product in seven different hospitals in Spain. HCV RNA was also investigated in two batches of the IVIG shared by some recipients. All the viral RNA detected were characterized with a line probe assay, restriction fragment length polymorphism analysis of the 5'-noncoding and core regions, and sequencing of the nonstructural 5 region. RESULTS: On the basis of both clinical and laboratory data, a relationship could be established between the IVIG therapy and the acquisition of the HCV infection by the recipients. Several HCV strains were detected among the recipients, with most of the recipients coming from the same hospital presenting with closely related strains. Moreover, an HCV strain almost identical to the main strain detected among the recipients was found in one batch of the IVIG that probably was shared by most of them. Follow-up studies and evaluation of low-avidity anti- HCV IgG suggested that both acute primary infections and reinfections were produced. In one case, direct evidence of reinfection by a different HCV strain was obtained. CONCLUSION: The results did not exclude the possibility that a second HCV strain associated with a further, unidentified batch of the IVIG could have contributed to this outbreak.     

GB virus type C infection in patients treated for childhood acute lymphoblastic leukemia

BACKGROUND: The purpose of this study was to determine the prevalence of GB virus type C (GBV-C) infection in subjects treated for childhood acute lymphoblastic leukemia (ALL) or non-Hodgkin's lymphoma. STUDY DESIGN AND METHODS: One hundred forty patients (82 males) aged 4 to 27 years (median, 11) diagnosed with ALL between 1976 and 1993, were prospectively followed for a median of 5 years (range, 0.1-17) after completion of therapy. Stored sera were tested for antibody to hepatitis C virus (HCV), HCV RNA, antibody to GBV-C E2 (anti-E2), and GBV-C RNA. RESULTS: Thirty-eight patients (27%) were exposed to GBV-C: 30 were positive for GBV-C RNA (mostly type 2) and 8 were positive for anti-E2. Anti-E2 and GBV-C RNA were mutually exclusive: 61 patients (43%) were positive for HCV RNA, 16 (11%) were coinfected with GBV-C and HCV. Alanine aminotransferase (ALT) levels were increased (>35 mU/mL) in 32 (23%) of 137: 3 of 20 who were positive for GBV-C and negative for HCV, 7 of 15 who were positive for GBV-C and HCV, 15 of 44 who were negative for GBV-C and positive for HCV, and 7 of 58 who were negative for GBV-C and HCV (p<0.001). Median ALT values were significantly higher in patients positive for GBV-C and HCV than in those who were positive for GBV-C and negative for HCV (35 vs. 13 mU/mL, p = 0.003). Thirty-one of 38 patients with GBV-C markers were retested: GBV-C RNA was lost in 16 of 30 tested, but 7 were still GBV-C RNA positive up to 50 months later, 3 tested positive for anti-E2 up to 27 months later, and 1 was positive for GBV-C RNA and anti-E2 26 months later, while 20 tested negative for both. CONCLUSION: GBV-C did not behave as a liver pathogen, because ALT alterations were unrelated to GBV-C status, but, rather, were related to HCV infection or coinfection. GBV-C RNA was frequently lost over a relatively short period, though in some cases, it was retained for a longer time. Anti-E2 rarely coexisted with GBV-C RNA and might be short-term.     

Investigation of an algorithm for anti HCV EIA reactivity in blood donor screening in Turkey in the absence of nucleic acid amplification screening

Purpose

In this study we aimed to propose an algorithm for initial anti HCV EIA reactive blood donations in Turkey where nucleic acid amplification tests are not yet obligatory for donor screening.

双抗原夹心ELISA检测抗HCV总抗体

目的建立检测抗HCV抗体的双抗原夹心ELISA法,评价其可行性。方法将与His或GST融合表达的HCV基因工程抗原,分别用作ELISA的包被和酶标记抗原,建立用于抗HCV总抗体检测的双抗原夹心ELISA。用此方法检测1 968份临床血清标本,并以间接ELISA(北京万泰试剂)与之对照;此外,用套式RT-PCR检测部分血清的HCV RNA。结果有1 761份血清2种ELISA检测均为阴性,有190份血清均为阳性,两种方法符合率为99.1%;有17份血清的检测结果不相符,间接法阳性而本法阴性的14份,其中HCV RT-PCR阳性1份;本法阳性而间接ELISA阴性的3份,其中RT-PCR阳性2份。双抗原夹心ELISA与间接ELISA的敏感性分别为99.48%、98.96%,特异性分别为99.94%、99.27%。结论新研制的检测抗HCV总抗体的双抗原夹心ELISA具有较高的敏感性和特异性,值得作进一步的深入研究。     

HCV RNA RT—PCR测定室间质量评价的研究

目的 建立国内ＨＣＶ ＲＮＡ ＲＴ－ＰＣＲ测定的室间质量评价系统。方法：室间质量评价研究工作采用的质控血清盘由１０支冻干质控血清组成，其中包括２份强阳性、２份弱阳性，２份阴性和一个５倍位比稀释系列（４份），将血清盘寄发给各类评实验室并要求各个实验室按照规定时间检测，回报结果，然后对结果进行统计分析。结果 多数参评实验室存在不同程度的假阳性和假阴性的问题。２份强阳性标本检出率均为８７．５％，弱阳性标     

人类免疫缺陷病毒/丙型肝炎病毒共感染对实验室诊断影响的研究

目的 研究人类免疫缺陷病毒（HIV）／丙型肝炎病毒（HCV）共感染对两种病毒感染实验室诊断的影响。方法 对300例经免疫印迹试验（WB）确认的HIV感染者，以酶免疫测定HCV抗体，测定CD4／CD8计数；其中197例测定病毒载量，逆转录聚合酶链反应检测HCV核酸，阳性样品限制性片段长度分析丙肝分型，比较共感染和非共感染组在各检测诊断指标的差别。结果 HCV诊断：HCV抗体阴性组中19．5％为核酸阳性。多元回归分析，HCV核酸阳性，1b＋2a混合感染，1b基因型的感染3个因素对HCV EIA检测的S／CO值的升高有独立的显著影响。HIV诊断：300例，共感染组和非共感染组HIV ELISA A〈3的样本比例分别为4／265，5／41，有P〈0．01的差异有统计学意义；A〉3的样品中CD4分组后按比例取77份比较WB条带，共感染组P55条带出现率显著高于非共感染组（16／30，12／47，P〈0．01）。结论 HIV免疫抑制可造成很高的HCV抗体假阴性率，该人群推荐HCV核酸定性检测。共感染对HIV ELISA检测的影响表现为强阳性比例的显著提高；对HIV WB各主要诊断条带未见影响，共感染组p55条带比例显著高可能提示病毒间免疫和分子水平相互作用。对HC VEIA-3的S／CO值有独立影响的因素包括HCV核酸阳性，1b＋2a混合感染，1b基因型。     

HGV/GB virus C transmission by blood components in patients undergoing open-heart surgery

BACKGROUND: To establish the rate of HGV/GB virus C (GBV-C) transmission by blood components in open-heart surgery patients. STUDY DESIGN AND METHODS: From 55 patients receiving blood components, sera were collected before and 2, 4, 6, 8, 10, 12, 16, 20, 26, and 32 weeks after heart surgery. Serum samples from patients and implicated blood donations were tested for HGV/GBV-C RNA by PCR. Recipients of RNA-positive blood components were also tested for the presence of E2 antibodies (E2Ab) by ELISA. RESULTS: Of 55 recipients, 18 received RNA-positive blood components. Of 14 recipients of RNA-positive blood components, who were negative for RNA or E2Ab before transfusion, 8 became RNA positive and one developed E2Ab after transfusion. Three recipients of RNA-positive blood components had E2Ab before transfusion, and none of these became RNA positive after transfusion. One of 18 recipients was RNA positive before and after transfusion. Of 55 recipients, 37 received RNA-negative blood components: 34 were RNA negative before and after transfusion. Of 37 recipients, 3 were RNA positive before and after transfusion. CONCLUSION: Of susceptible patients, 64 percent became infected with HGV/GVC-C when transfused with RNA-positive blood components. E2Ab-positive patients were protected against HGV/GBV-C infection.