肝细胞癌患者GB病毒C感染状况及其相关性研究

为了解ＧＢ病毒Ｃ（ＧＢＶ－Ｃ）感染与肝细胞癌（下称肝癌）之间可能的相关性,采用一步法逆转录聚合酶链反应（ＲＴ－ＰＣＲ）对１２４ 例肝癌患者进行了ＧＢＶ－ＣＲＮＡ 检测。结果：ＧＢＶ－ＣＲＮＡ 总检出率高达２６．６％ （３３／１２４）,３３ 例ＧＢＶ－ＣＲＮＡ 检出阳性者中,９０．９％ 合并有ＨＢＶ、ＨＣＶ感染;３３ 例非Ｂ非Ｃ型肝癌中,ＧＢＶ－ＣＲＮＡ检出率为９．１％ （３／３３）。３３ 例ＧＢＶ－Ｃ感染阳性肝癌患者中,９０．０％ （３０／３３）有既往输血史,高于ＧＢＶ－Ｃ感染阴性组（４８／９１）。上述结果提示,我国肝癌患者中ＧＢＶ－Ｃ感染的检出率高于美、欧、日等地同类患者的检出率,其在肝细胞癌及输血后肝炎发生中的病因学意义值得注意。     

乙,丙型肝炎病毒单纯和重叠感染的病毒学和血清学分析

探讨乙型肝炎病毒（ＨＢＶ）与丙型肝炎病毒（ＨＣＶ）重叠感染对ＨＢＶ或ＨＣＶ复制的影响。方法检测１２４例各型肝炎血清ＨＢＶＤＮＡ、ＨＣＶＲＮＡ、抗ＨＣＶ和ＨＢＶ５项标志，对比分析ＨＢＶ或ＨＣＶ单纯感染及两者重叠感染时血清ＨＢＶ复制标志［ＨＢＶＤＮＡ及（或）ＨＢｅＡｇ］或ＨＣＶ复制标志（ＨＣＶＲＮＡ）的检出率。结果ＨＣＶ单纯感染组ＨＣＶＲＮＡ检出率８３．３３％（２５／３０），高于ＨＣＶ与ＨＢＶ重叠感染组５５．５６％（Ｐ＜０．０５），临床类型分析显示这种差异发生在慢性肝炎组；ＨＢＶ单纯感染组复制标志检出率为４１．８６％（１８／４３），重叠感染组ＨＢＶ复制标志检出率为３０．５６％（１１／３６），两组差异无显著性。结论ＨＢＶ与ＨＣＶ重叠感染时ＨＣＶ复制受到抑制，ＨＢＶ复制标志无明显影响     

RIBA在慢性丙型病毒性肝炎中的应用价值

采用含ＨＣＶ三种抗原（Ｃ２２，Ｃ３３Ｃ，Ｃ１９０）的重级免疫印迹试验（ＲＩＢＡ）检测了７１例抗－ＨＣＶ阳性一（ＲＩＡ－２法）的慢性非甲非乙型肝炎患者血清并与ＨＣＶＲＮＡ结果进行比较。ＲＩＢＡ阳性５６例，可疑８例，对应ＨＣＶＲＮＡ检出率分别为９４．６４％（５３／５６）和３７．５％（３／８），ＲＩＢＡ阴性的７例均未检出ＨＣＶＲＮＡ。ＨＣＶＲＮＡ阳性病例中ＲＩＢＡ多为阳性，可疑仅占５．３６％（３／５６０     

丙型肝炎患者尿液特异标志物的检测及其临床意义

对５４例丙型肝炎患者尿液特异指标检测，结果：抗－ＨＣＶ阳性５／５４例（９．２％）。ＨＣＶＲＮＡ阳性９／５４例（１６．７％）。急性ＨＣＶ感染者，尿中抗－ＨＣＶ检出率为２／６例（３３％），较慢性者３／３８例（７．９％）为高（Ｐ＜０．０５）。尿液ＨＣＶＲＮＡ全部出现在未经α－干扰素治疗的慢性患者中，故可用作判断干扰素疗效的指标之一。本文在尿中检出ＨＣＶＲＮＡ，提示ＨＣＶ可经尿液污染环境。     

人肝癌组织中黄曲霉毒素和肝炎病毒基因检测分析

以肝细胞场（ＨＣＣ）患者手术切除标本为研究材料，采用高效液相色谱荧光法检测组织中的黄曲霉毒素Ｂ１（ＡＦＢ１），嵌套式多聚酶链反应技术检测组织中的ＨＢＶＤＮＡ和ＨＣＶＲＮＡ。在６９．２％（１８／２６例）的ＨＣＣ患者中检出了３．６５－８９．０６ｎｇ不等的ＡＦＢ１／ｇ肝组织；ＨＢＶＤＮＡ和ＨＣＶＲＮＡ检出率分别为９０．５％（５７／６３例）和１２．７％（８／６３例）。其中，ＡＦＢ１和ＨＢＶＤＮＡ重叠检出率为６１．５％（１６／２６例），ＨＢＶＤＮＡ和ＨＣＶＲＮＡ重叠检出率为７．９％（５／６３例），４例ＨＣＣ患者重叠检出了ＡＦＢ１、ＨＢＶＤＮＡ和ＨＣＶＲＮＡ；同步检测对照组５例，无１例出现这种情况。结果表明，上述３种因素均与人类肝癌密切相关，并且可能存在协同致癌效应。     

慢性肝病患者庚肝病毒RNA检测分析

应用逆转录酶链反应技术（ＲＴ－ＰＣＲ）对９６例慢性肝病患者进行ＨＧＶ ＲＮＡ检测,结果：慢性肝炎９例,肝硬化５６例,肝癌３１例中ＨＧＶ ＲＮＡ阳性分别为１例,６例,３例,总检出率为１０．４２％（１０／９６）在ＨＢＶ感染６３例,ＨＣＶ感染,８例及ＨＢＶ＋ＨＣＶ重叠感染１１例中,ＨＧＶ ＲＮＡ阳性分别为８例,１例和１例,在慢性肝病中ＨＧＶ ＲＮＡ阳性与阴性组间,ＡＳＴ,ＡＬＴ水平无显著性差异（Ｐ〉０．     

肝硬化,肝癌患者乙型及丙型肝炎病毒的检测

对４１例肝硬化病人，４０例肝癌病人及４１例非肝炎、非肝硬化及非肝癌病人（对照组）因清乙肝５项（ＨＢｓＡｇ，抗ＨＢｓ、抗ＨＢｃ、ＨＢｅＡｇ、抗ＨＢｅ）、血清抗ＨＣＶ及血清ＨＣＶＲＮＡ检测。结果：肝硬化组、肝癌组和对照组乙肝５项检出率分别为：９２．７％９５．０％６ ３１．７％；抗ＨＣＶ检出率分别为３１．７％１２．５、和９．７％；ＨＣＶＲＮＡ检 分别为：３６．６％、２７．５％和１４．６％。通过比较分析认     

应用聚合酶链反应进一步探讨原发性肝癌与HBV,HCV感染的关系

应用聚合酶链反应（ＰＣＲ）检测原发性肝癌（ＰＨＣ）及肝硬化（ＬＣ）患者血中ＨＢＶ－ＤＮＡ及ＨＣＶ－ＲＮＡ，同时检测血中乙肝５项标志及抗－ＨＣＶ。结果５６例ＰＨＣ中，ＨＢＶ５项标志阳性者４３例（７６．７８％），ＨＢＶ－ＤＮＡ阳性者４２例（７５．００％），两种方法联合应用阳性率为４９例（８７．５％）；其中３４例检测ＨＣＡ－ＲＮＡ阳性者８例（２３．５３％）。６２例ＬＣ中，ＨＢＶ５项标志阳性者３５例（５６．４５％），ＨＢＶ－ＤＮＡ阳性者３８例（６１．２９％），二者联合应用阳性数为４５例（７２．５８％），其中３４例检测ＨＣＶ－ＲＮＡ阳性数为９例（２３．６８％）。ＰＨＣ组ＨＢＶ感染率显著高于ＬＣ组（Ｐ＜０．０５），ＨＣＶ感染率两组差异无显著性（Ｐ＜０．０５）。并对ＨＢＶ、ＨＣＶ感染与ＬＣ、ＰＨＣ的发生发展作进一步讨论。     

荧光定量PCR技术在检测HCV—RNA中的应用

目的 评价荧光定量ＰＣＲ技术测定ＨＣＶ－ＲＮＡ水平的价值,探讨ＰＢＭＣ中检出ＨＣＶ－ＲＮＡ的意义。方法 采用荧光定量ＫＲ及ＲＴ－ＰＣＲ技术同时检测８７例血液透析患者的血清和淋巴细胞中ＨＣＶ－ＲＮＡ。结果血清、淋巴细胞中ＨＣＶ－ＲＮＡ的荧光定量ＰＣＲ检出率（４９．４％、６９．０％）,分别高于相应标本中ＨＣＶ－ＲＮＡ的ＲＴ－ＰＣＲ阳性率（３３．３％、３５．６％）（Ｐ ＜０．０５）。血清、淋巴细胞中同时检出ＨＣＶ－ＲＮＡ的一致率,荧光定量ＰＣＲ法（３１．０％）显著高于ＲＴ－ＰＣＲ定性法（６．９％）（Ｐ＜０．００１）． 荧光定量ＰＣＲ法,淋巴细胞中ＨＣＶ－ＲＮＡ检出率高于血清（Ｐ＜０．０１）。结论 与 ＲＴ－ＰＣＲ相比,荧光定量ＰＣＲ技术检测 ＨＣＶ－ＲＮＡ敏感性较高,对ＰＢＭＣ内ＨＣＶ－ＲＮＡ检测有利于提高ＨＣＶ检测的阳性率。     

肝癌组织中丙型肝炎病毒RNA的研究

对３０例原发性肝癌患者手术切除的癌及癌旁组织（共３８份标本）用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）扩增丙型肝炎病毒（ＨＣＶ）５＇端非编码区（５＇－ＮＣ区）ＲＮＡ。结果，１０例（３３．３％）肝癌患者被ＨＣＶ感染，２３例（７６．７％）患者被乙型肝炎病毒（ＨＢＶ）感染。用酶切分析可见，肝癌组织中的ＨＣＶ是ＩＩ、ＩＩＩ型。对部分标本用ＰＣＲ直接测定核苷酸序列，ＨＣＶ５＇－ＮＣ区相当保守，差异无显著意义。因１０例ＨＣＶ感染的肝癌患者中８例同时有ＨＢＶ感染，因此可能ＨＣＶ及ＨＢＶ构成的双重慢性感染是导致肝癌发生的因素之一。     

血液透析患者庚型肝炎病毒感染的研究

目的探讨血透患者庚型肝炎病毒(HGV)感染情况及其感染相关因素。方法采用ELISA法和RT—PCR法检测264例血透患者抗HGV和HGV—RNA，并同时检测抗HCV、HCV—RNA、HBsAg和抗HBc。结果血透人群HGV感染率为13．6％．其中，36例HGV阳性患者有21例与其它肝炎病毒合并感染(58．3％)，6例(16．7％)与HBsAg，6例(16．7％)与抗HCV／HCV—RNA，9例(25．0％)与抗HBc／HBsAg／抗HCV。HGV—RNA与HBV、HCV合并发生率为66．7％。HGV感染以女性人群多见，与年龄、输血史、输血次数、透析时间、HBV感染及HCV感染无相关性，感染者肝功能生化指标无异常改变。结论血透人群有较高的HGV感染率，其中多数感染与HBV、HCV合并发生。     

原发性肝癌78例血清HCVRNA和抗HCV的观察

用逆转录－聚合酶链反应和酶联免疫吸附试验检测了７８例原发性肝癌患者清丙型肝炎病毒（ＨＣＶ）ＲＮＡ、抗ＨＣＶ和乙型肝炎病毒（ＨＢＶ）免疫学标物。     

Differential influence of different hepatitis viruses on quality of life in HIV positive patients

HIV-infected individuals are frequently co-infected with different hepatitis viruses. HCV has been associated with impaired quality of life in non-HIV infected patients. Little is known concerning the quality of life in HIV-infected individuals in relation to the different viral co-infections. - Patients and Methods: We investigated 250 patients who have answered HIV-SELT and EuroQoL (EQ-5D) questionnaires assessing quality of life. Data on HBsAg, anti-HBc, anti-HCV, and GBV-C-RNA were available for 191, 188, 189, 98 patients, respectively. HCV-RNA was tested in 33 of 35 anti-HCV positive patients. - Results: There was no difference in quality of life in relation to active or past HBV-infection defined by HBsAg (n = 15) and anti-HBc in the absence of HBsAg (n = 84), respectively, for both overall HIV-SELT (p = 0.66, and p = 0.43, respectively) and visual EQ-5D (p = 0.93 and p = 0.64, respectively). However, anti-HCV positivity (n = 35) was associated with significantly impaired quality of life (HIV-SELT overall p<0.001). Importantly, no difference was found in relation to HCV-viraemia in anti-HCV positive patients (p = 0.77). In multivariate analysis anti-HCV positivity, employment status, HIV viral load and GBV-C were relevant to quality of life, with GBV-C being beneficial and HCV being negative. - Conclusions: While HBV seems to play no role concerning quality of live in HIV-infected patients, the flavi-viruses HCV and GBV-C display opposing influence on quality of life. As quality of life was similarly impaired in HCV-viraemic and HCV-non-viraemic anti-HCV positive patients but better in GBV-C viraemic patients, this should be taken into account in the indication case of planned interferon therapy.     

肝细胞癌患者血清与癌组织中抗HCV及HCVRNA的检测

８８例经病理确诊的肝细胞癌患者，用ＥＬＩＳＡ法检测其血清抗ＨＣＶ与ＨＢＶ－Ｍ，阳性率分别为１４．７７％和９０．９１％。用ＰＣＲ法检测发现血清ＨＣＶＲＮＡ阳性１２例（１３．６４％）；１１例血清ＨＢＶＤＮＡ阳性病人的肝癌组织中８例ＨＢＶＤＮＡ阳性；１例肝癌组织中同时检出ＨＣＶＲＮＡ与ＨＢＶＤＮＡ。结果表明，本组肝癌的发生主要与ＨＢＶ感染有关，而与ＨＣＶ感染也可能有一定关系。     

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     

Comparative assay of Hepatitis B and C virus infection markers by different assay kits

In order to compare sensitivity of EIA and RIA assay kits for hepatitis B and C virus(HBV and HCV,respectively)infection markers,100 serum samples in total wre collected form 50 adult women each in urban and rual areas in northeast China.The number of positive cases to the three infection markers on HBV(i.e..,HBsAg^ ,anti-HBs^ ,and anti-HBc^ )and the one on HVC (anti-HCV)were examined in two laboratories,i.e.,in Laboratory A with EIA kits produced in China and in Laboratory B with RIA kits.HCV infection positivity(anti-HCV^ )was examined by EIA kits in both laboratories,but from diffeent sources in and outside of China,respectively.The assay in Laboratory A gave 2 HBsAg^ cases out of the 100 cases examined,whereas there were 9 positive cases in Labortory B,In contrast,19 cases were positive to anti-HCV when examined in Laborator A,and there were 3 cases in Laboratory B.Thus,the kits used in Laboratory A gave fewer HBsAg^ and more anti-HCV^ cases than the kits used in Laboratory B.The prevlence of antiHBs^ or anti-HBc^ and cases did not differ when assayed in the two laboratories with EIA and RIA kts,respectively,The agreement of positive and negative findings between the two set of testing were 93%,93%,93%,86%and 82% for HBsAg,anti-HBs,anti-HBc,BHV(i.e.,either positive to anyone of the three markers or negative to all three markers),and anti-HCV,respectively.The implication of the observation on epidemiology on HBV and HCV infection prevalence was discussed.     

PCR检测丙型肝炎病毒感染各高危人群

用ＰＣＲ方法克隆了中国丙型肝炎病毒（ＨＣＶ）感染者的５’端非编码区（５’－ｎｏｎｃｏｄｉｎｇｒｅｇｉｏｎ，５’－ＮＣＲ）序列，并根据此序列合成引物检测我国ＨＣＶ各高危感染人群的病毒复制情况，结果显示ＨＣＶＲＮＡ在慢性丙型肝炎患者的血清标中的阳性率为８１．８％（９／１１），在ＨＣＶ抗体阳性的血液透析患者血清标本中的阳性率为６３．６％（７／１１），在ＨＣＶ抗体阳性的献血员血清标中的阳性率为２０％（４／     

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.     

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