High prevalence of hepatitis C virus-ribonucleic acid positivity in anti-hepatitis C virus negative renal transplant patients

Background

Hepatitis C virus (HCV) infection is common in renal transplant (RT) patients. Some of these patients remain anti-HCV negative despite presence of infection and these are identified by a positive HCV-ribonucleic acid (RNA) test.

Methods

We studied 404 RT patients for prevalence of HCV-RNA positivity in anti-HCV negative patients. Serum was tested for presence of anti-HCV antibodies using a third generation HCV micro-ELISA (enzyme-linked immunosorbent assay) test, which utilises a combination of HCV structural and nonstructural antigens. The RNA was extracted from patient serum for HCV viral quantification using Quiagen Ql Amp Viral RNA mini extraction kit. The HCV-RNA viral load was performed on Corbet Rotor Gene 3000 thermocycler using Taqman principle.

Results

About 308 patients were anti-HCV negative and 96 were anti-HCV positive, resulting in prevalence of overt HCV infection of 23.7%. A total of 130 anti-HCV negative patients tested positive for HCV-RNA making a prevalence of occult HCV infection of 42.2%. There was no significant difference in the rate of overt or occult HCV infection between males and females. Patients with HCV infection (whether overt or occult) had received more number of dialysis sessions (62.5 vs 32.2) and blood transfusions (2.78 vs 1.99) when compared to those without HCV infection (P=0.001). The mean duration on dialysis was also longer (8.15 months vs 4.53 months) in patients with HCV infection (P= 0.0001).

Conclusion

A direct test for HCV viraemia is important to accurately determine the epidemiology of HCV infection in RT patients who remain anti-HCV negative despite harbouring active HCV infection.     

High prevalence of hepatitis C virus-ribonucleic acid positivity in anti-hepatitis C virus negative renal transplant patients

Background

Hepatitis C virus (HCV) infection is common in renal transplant (RT) patients. Some of these patients remain anti-HCV negative despite presence of infection and these are identified by a positive HCV-ribonucleic acid (RNA) test.

Methods

We studied 404 RT patients for prevalence of HCV-RNA positivity in anti-HCV negative patients. Serum was tested for presence of anti-HCV antibodies using a third generation HCV micro-ELISA (enzyme-linked immunosorbent assay) test, which utilises a combination of HCV structural and nonstructural antigens. The RNA was extracted from patient serum for HCV viral quantification using Quiagen Ql Amp Viral RNA mini extraction kit. The HCV-RNA viral load was performed on Corbet Rotor Gene 3000 thermocycler using Taqman principle.

Results

About 308 patients were anti-HCV negative and 96 were anti-HCV positive, resulting in prevalence of overt HCV infection of 23.7%. A total of 130 anti-HCV negative patients tested positive for HCV-RNA making a prevalence of occult HCV infection of 42.2%. There was no significant difference in the rate of overt or occult HCV infection between males and females. Patients with HCV infection (whether overt or occult) had received more number of dialysis sessions (62.5 vs 32.2) and blood transfusions (2.78 vs 1.99) when compared to those without HCV infection (P=0.001). The mean duration on dialysis was also longer (8.15 months vs 4.53 months) in patients with HCV infection (P= 0.0001).

Conclusion

A direct test for HCV viraemia is important to accurately determine the epidemiology of HCV infection in RT patients who remain anti-HCV negative despite harbouring active HCV infection.Key Words: hepatitis C, real time PCR, renal replacement therapy     

血液透析患者中TT病毒感染的检测及序列分析

目的：研究血液透析患中TT病毒（TTV）的感染状况及其致病性。方法：针对病毒基因保守区设计引物,采用套式PCR方法对69例血液透析患血清标本进行TTVDFNA检测及序列分析。并同时检测患血清丙氨酸转氨酶（ALT）及丙型肝炎病毒抗体（抗-HCV）。结果：患血清TTVDNA总阳性率为27.5%。对其中1例PCR扩增产物测序。结果与其他TVV分离株如GH1、TA278、TTVCHN1和TTVCHN2的核苷酸序列同源性为89%-100%,推测的氨基酸序列同源性为87%-100%。抗-HCV阳性与阴性患的TTV DNA阳性率无明显差异。所有患均未见ALT明显升高。结论：血液透析患存在较严重的TTV感染与HCV感染无明显相关性。未发现TTV感染导致ALT明显升高的证据。     

丙型肝炎病毒核心抗原的检测及其临床意义

目的 探讨HCV 核心抗原在HCV 感染诊断和治疗中的作用.方法 丙肝核心总抗原检测采用酶联免疫法;丙型肝炎抗体检测采用第三代酶联免疫法.所有血清标本均进行HCV-RNA 和肝功能检测.丙肝核酸定量采用PCR- 荧光探针法;肝功能检测采用紫外连读检测法.结果 90 例中,HCV-cAg 检测阳性率为42.22%(38/90).HCV-cAg 阳性组HCV-RNA 阳性率100%(38/38),显著高于HCV-cAg 阴性组的71.15%(37/52).阳性组丙氨酸氨基转移酶(alanine aminotransferase,ALT)异常率63.16%(24/38),显著高于HCV-cAg 阴性组的40.38%(21/52),两组间比较(P<0.05).HCV-cAg 阴性组中59.62% (31/52)的患者ALT 在正常范围内(ALT<40U/L),显著高于HCV-cAg 阳性组的36.84%(14/38) ;而HCV-cAg 阳性组中36.84%(14/38)ALT 值均>100U/L,明显高于HCV-cAg 阴性组的9.62%(5/52).两组间比较(P<0.05).丙肝核心抗原HCV-cAg 和HCVRNA有很好的正相关性,HCV-cAg 与ALT 水平也有一定相关性,可以反映肝功损害程度.结论 丙肝核心抗原HCV-cAg是慢性丙肝感染良好的监测指标.     

丙型肝炎患者唾液和尿HCV－RNA检测及其意义

采用套式聚合酶链反应检测丙肝患者血清，唾液和尿各５２份。结果血清ＨＣＶ－ＲＮＡ阳性３９例，２３例分别在尿和唾液中检出ＨＣＶ－ＲＮＡ；１３份血清ＨＣＶ－ＲＮＡ阴性者中，尿和唾液均未检出ＨＣＶ－ＲＮＡ，有非常显著差异（Ｐ＝０．０００１）。血清ＨＣＶ－ＲＮＡ阳性者分ＡＬＴ正常组和异常组，尿和唾液ＨＣＶ－ＲＮＡ阳检率分别为１８％和７５％，亦有非常显著差异（Ｐ＝０．００１）。对照组１３例正常人血清、唾液和尿ＨＣＶ－ＲＮＡ均为阴性。表明丙肝患者尿和唾液中能否检出ＨＣＶ－ＲＮＡ与病毒血症及ＡＬＴ有关，提示丙肝患者的尿和唾液可能为ＨＣＶ感染的传播媒介。     

Hepatitis C virus infection in hemodialysis patients]

To study whether HCV infection is present in dialysis patients and the status of antibody responses to HCV viremia, 16 patients undergoing hemodialysis were assayed for anti-HCV by ELISA and RIBA. A high incidence of anti-HCV (13/16) was found. All the 16 patients were studied for HCV viremia by 'nested' PCR and 10 were HCV RNA positive. Altogether 14 were anti-HCV and/or HCV RNA positive, which indicates that the incidence of HCV infection is high in hemodialysis patients, at least in some units. Cloning, sequencing of the PCR products showed high homology among isolates, with 91% homology to that of Japanese strains. It is important to study anti-HCV/HCV RNA in dialysis patients.     

血液透析患者丙型肝炎病毒感染的临床研究

为了探讨血透患者丙型肝炎病毒（ＨＣＶ）的感染情况及其影响因素，采用第二代ＥＬＩＳＡ法和ｎｓｅｔ－ＰＣＲ分别测定１４０例血透患者血清中ＨＣＶ和ＨＣＶ ＲＮＡ。结果：抗ＨＣＶ和ＨＣＶ ＲＮＡ的阳性率在血透组和非血透组之间差异非常显著，且随着血透时间延长而增高，５个以上患者达９４．６％；ＨＣＶ感染率与否输过血无关；采取严格隔离透析、透析器复用等预防措施可降低ＨＣＶ感染率。结论：血透明显增加丙型肝炎的感染，且与输血无关，预防措施有助于降低ＨＣＶ在血透患者中的发生率。     

维持性血液透析患者的丙型病毒性肝炎感染率

目的　评价血液透析(血透)患者丙型肝炎病毒(HCV)感染状况、HCV阳转率及危险因素。方法　采用第2代酶链免疫吸附试验(ELISA2,和逆转录聚合酶链反应(RTPCR)检测62例血透患者血清抗HCV和HCVRNA,采用ELISA2随访1995年6月～1996年12月间199例血透患者HCV阳转情况。结果　62例患者中,抗HCVIgM阳性27例(43.6%),抗HCVIgG阳性29例(46.8%),HCVRNA阳性34例(54.8%),3项至少1项阳性37例(59.7%)。HCV阳性组与阴性组相比,两组在性别、年龄、肾功能、HBV标志、EPO应用、CAPD史无明显差异,而在平均透时间、输血史、ALT异常、肾移植史方面有显著性差异。1995年6月抗HCV阳性率为58.7%,每隔半年的阳性率分别为53.7%、54.8%和50.0%。在18个月随访期间,31例阳转;156例随访1～6月,阳转14例(8.9%),67例随访7～12月,15例(22.4%)阳转,6例随访13～18月,2例(33.3%)阳转;2例阳转并转为肝硬化。结论　血透患者中HCV感染严重,输血为血透患者感染HCV的主要途径,但也存在医源性传播的可能。     

慢性丙型肝炎患者HCV不同基因片段抗体应答与脂代谢关系

目的:探讨慢性丙肝患者HCV感染与脂质代谢的关系及特点。方法:用酶联免疫法检测实验组慢性丙肝患者HCV不同基因片段抗体,并以羊抗人Apo-B血清分离其Apo-B,用PCR法检测Apo-B中HCV-RNA,同时作血脂监测。另设健康人和乙肝病例对照。结果:实验组病例HCV不同基因片段抗体呈现6种分布模式,其血清中均检出HCV-RNA较高载量,相应的Apo-B中(除1例阴性外)均有HCV-RNA表达,血清LDL、Apo-B等水平值显著低于对照组。两对照组血清及Apo-B中无HCV-RNA表达,且不同基因片段抗体阴性。结论:慢性丙肝患者HCV不同基因片段抗体应答与HCV复制密切相关,与Apo-B携带密切相关;其HCV-C和HCV-NS4、HCV-NS5与不同组分载脂蛋白相互作用,可能共同干扰脂质代谢。     

抗-HCV和HCV-RNA检测及其与ALT的相关性分析

目的探讨化学免疫发光法(CLIA)定量检测抗-HCV和FQ-PCR法检测HCV-RNA含量与丙氨酸氨基转移酶(ALT)水平的相关性。方法用CLIA定量筛选抗-HCV阳性的100例病人标本,以荧光定量PCR法检测HCV-RNA含量和酶速率法检测ALT浓度水平,并对所得数据进行统计分析。结果在100份抗-HCV阳性标本中,检出HCV-RNA阳性者76例,阳性率为76%。随着抗-HCV的S/CO值增高,HCV-RNA检出率增高较明显;ALT水平与HCV-RNA含量无显著相关性(P>0.05),但ALT异常率与HCV-RNA含量呈正相关。结论在HCV诊断与疗效观察中,血清抗HCV、HCV-RNA和ALT指标各有利弊,3者有机结合能正确诊断和预测肝脏损伤及评价疗效。     

HCV-cAg与HCV-Ab、HCV-RNA及肝功能之间的关系探讨

目的探讨丙型肝炎病毒核心抗原（hepatitis C virus core antigen,HCV-cAg）与丙型肝炎病毒RNA（hepatitis C virus RNA,HCV-RNA）关系以及丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、谷氨酰转肽酶（GGT）水平与HCV-RNA病毒载量之间的相关性。方法选取78例HCV抗体检测阳性的丙型肝炎或疑似丙型肝炎病人作为观察组,另选取同期健康体检者20名作为对照组。采用ELISA法和荧光定量PCR法等对2组血清样本进行HCV-cAg、HCV-RNA和ALT、AST、GGT水平测定,并比较各参数之间的关系。结果观察组HCV-cAg阳性检出率为41.0%（32/78）,HCV-RNA阳性检出率为53.8%（42/78）,差异有统计学意义（P < 0.01）,且二者之间具有较好一致性（κ=0.747）。观察组ALT、AST和GGT水平均明显高于对照组（P < 0.01）,但与HCV-RNA病毒载量均无明显相关性（P>0.05）。结论HCV-cAg和HCV-RNA在检测丙型肝炎方面有较好一致性和相关性,可为丙型肝炎的临床诊断提供有价值的依据。     

Hepatitis C Virus Infection in Haemodialysis Patients: “Wolf in Sheep's Clothing”

Background

170 million people are infected with the Hepatitis C virus (HCV) around the world. Approximately 50%-70% patients infected with HCV develop chronic liver disease. Haemodialysis patients constitute an especially important group with high HCV prevalence. Outbreaks of HCV infection in dialysis units have been documented. Detection of anti-HCV antibodies is a convenient and conventional mode of documentation. However, in this group, it has it''s own caveats.

Methods

48 patients who had undergone or were on haemodialysis (HD) and had undergone a minimum of 15 dialysis sittings were studied. HCV infection was documented both by anti-HCV antibody detection and HCV RNA testing. A comparative evaluation of results by both tests was done.

Results

Out of a total of 48 patients, HCV RNA was detected in 38 (79.16%) and anti-HCV antibodies in 13(27.07%). Out of 48 patients 10(20.83%) were negative for both parameters. 22.91% (11/48) of patients were positive for both HCV RNA and anti-HCV antibody. 56.25% (27/48) were HCV RNA positive but anti-HCV antibodies were not detectable in their sera. 2 patients (04.16%) had a positive anti-HCV antibody status despite HCV RNA being negative. In 20.83% (10/48) both parameters were undetectable.

Conclusion

Chronic liver disease (CLD), particularly due to HCV infection, is a major complication amongst haemodialysis (HD) patients. Without reliable assays for antigenemia and the inability of antibody tests to define viremia in all cases, the detection of viral nucleic acid is necessary for diagnosis of active HCV infection.Key Words: Hepatitis C virus, Haemodialysis     

烧伤病人输血后丙肝发生情况报告

采用酶联免疫反应（ＥＬＩＳＡ）及聚合酶反应（ＰＣＲ）方法，对２０例烧伤病人血清丙型肝炎病毒抗体（抗－ＨＣＶ）及丙型肝炎病毒（ＨＣＶ）ＲＮＡ进行了检测。结果：入院时，输血组及非输血组抗－ＨＣＶ及ＨＣＶＲＮＡ检测均为阴性。一个月后。输血组１０例病人中发现抗－ＨＣＶ阳性５例、ＨＣＶＲＮＡ阳性４例；而非输血组抗－ＨＣＶ及ＨＣＶＲＮＡ检测均为阴性。说明输血是烧伤病人感染ＨＣＶ的重要途径。本还对烧伤病人输血     

122例艾滋病患者合并乙肝病毒 丙肝病毒感染的临床流行病学研究

目的 研究在抗逆转录病毒药物治疗(HAART)状态下HIV/AIDS患者合并乙肝病毒(HBV)、丙肝病毒(HCV)的流行状况以及相关血清生化和病毒学特征.方法 对122例正在接受HAART治疗的HIV/AIDS患者采集血标本,使用酶免法(ME-IA)检测乙肝五项指标(HBs Ag、抗-HBs、HBe Ag、抗-HBe、抗-HBc)、肝功能,应用酶联免疫吸附法(ELISA)检测丙型肝炎病毒抗体,运用RT-PCR法对丙肝病毒抗体阳性标本以及表面抗原和/或核心抗体阳性标本分别进行HCV-RNA以及HBV-DNA测定.以43名单纯HCV感染者作为对照组进行研究.结果 122名 HIV/AIDS患者,乙肝五项指标全阴性97例(78.86％),表面抗原阳性3例(2.46％),表面抗体阳性者总计19例(15.57％),单纯核心抗体阳性者3例(2.46％).HCV 抗体阳性84例(68.85％),84例HCV抗体阳性标本中HCV-RNA 阳性58例(69.05％),HIV/HCV/HBV合并感染者为0.HIV/HCV合并感染组 HCV-RNA 水平与感染者性别,CD4 +细胞数水平无明显差异(P＞0.05);HIV/HCV合并感染组与单纯HCV感染组 HCV-RNA水平、肝功能异常率无明显差异(P＞0.05).结论 HIV感染者HBsAg阳性率低于普通人群,HIV/HCV合并感染的比例显著高于普通人群.HIV/HCV合并感染与单纯HCV感染者血清生化及病毒学改变相似.     

HBV与HCV重叠感染的研究

应用ＰＣＲ法和套式ＰＣＲ法对深圳地区４４例ＨＢＶ血清学标志物阳性者进行ＨＣＶ－ＲＮＡ检测，并同时作ＨＢＶ－ＤＮＡ检测，以了解ＨＢＶ与ＨＣＶ重叠的感染情况，同时观察其血清ＡＬＴ的情况。结果发现，４４例患者中ＨＢＶ－ＤＮＡ阳性１２例（２７．２７％）；ＨＣＶ－ＲＮＡ阳性２２例（５０．００％），抗ＨＣＶ阳性２１例（４７．７３％）．表明ＨＢＶ与ＨＣＶ的重叠感染率较高，应引起人们的重视。     

血透患者血清庚型肝炎病毒RNA及抗体检测

目的:了解血透患者庚型肝炎病毒(HGV)感染率及与其他肝炎病毒重叠感染的情况。方法:收集57份血透患者血清标本,根据HGV RNA保守片段5′非编码区序列,设计了两对引物,采用逆转录巢式PCR扩增HGV RNA,以及EIA法检测HGV IgG抗体。同时还检测了HBV DNA及HCV IgG抗体。结果:57份标本中,4份HGV RNA阳性(7.0％),2份HGV IgG抗体阳性(3.5％)。HGV RNA与HGV IgG抗体呈“分离”现象。4份HGV RNA阳性标本中2份为HBV DNA阳性,2份为HCV IgG抗体阳性;其中1份标本HBV DNA和HCV IgG抗体均为阳性。结论:血透患者HGV感染率高于健康献血员和正常人群,HGV可与HBV和HCV重叠感染。     

慢性丙型病毒性肝炎患者相关自身抗体、血清HCV RNA水平及AST/ALT比值的关系探讨

目的对比分析丙型肝炎患者血清自身抗体、HCV RNA水平和AST/ALT比值,以进一步探讨丙型病毒性肝炎的发病机理。方法采用间接免疫荧光法测定抗核抗体（ANA）、抗平滑肌抗体（SMA）、抗线粒体抗体（AMA）和抗肝肾微粒体抗体（LKM）,采用核酸扩增（PCR）荧光定量技术检测标本HCV RNA含量,并同时检测肝功能指标丙氨酸氨基转移酶（ALT）、门冬氨酸氨基转移酶（AST）。结果HCV RNA阳性组和HCV RNA阴性组自身抗体检出率差异无统计学意义（P〉0.05） 自身抗体阳性组和自身抗体阴性组中AST/ALT比值≥1检出率有显著性差异（P〈0.01） HCV RNA阳性组和HCVRNA阴性组AST/ALT比值≥1检出率有显著性差异（P〈0.01）。结论病毒的直接作用和自身免疫是HCV损伤机体的两个重要因素。     

维持性血液透析患者合并感染的临床分析

目的：分析维持性血液透析（MHD）患者合并感染的特点和相关因素。方法：回顾48例MHD患者的感染部位、病原菌种类、免疫功能、营养状况、肾功能、原发病因等与感染的相关性。结果：感染组24例患者共发生感染33次,肺部感染和静脉导管感染最常见。行细菌培养29次,培养阳性20次,病原学检查以革兰氏阴性杆菌最常见（占34.5%）。感染组血红蛋白、血浆白蛋白明显降低。糖尿病和系统性红斑狼疮（SLE）患者感染发生率高。结论：MHD患者感染发生率高,积极改善贫血和营养状况,尽可能减少侵入性操作,有助于感染的防控。     

维持血液透析病人乙型丙型肝炎病毒感染的状况分析

目的探讨长期维持性血液透析(HD)患者感染乙丙型肝炎病毒感染情况。方法用ELISA和PCR法检测242例血液透析患者血清中HBVM、抗HCV和HCV-RNA。结果血透患者HCV阳性率33.5%,明显高于对照组(P<0.01),血透患者输血组明显高于非输血组(P<0.01),血液透析患者HBVM阳性率64.7%,输血组与非输血组的乙肝病毒感染无明显差别(P>0.05)。结论血透易感乙肝、丙肝病毒,输血是引起血透患者丙肝病毒感染的最主要原因。接受异体输血的次数和数量越多,感染的机率越大,阳性检出率则越高。