丙肝抗体、丙肝病毒RNA、白蛋白及谷丙转氨酶在丙肝诊断中的应用

目的:探讨丙肝抗体、丙肝病毒(HCV)RNA、白蛋白(ALB)及谷丙转氨酶(ALT)在丙肝诊断中的应用。方法:采用ELISA法测定丙肝抗体,实时荧光定量PCR检测HCV-RNA含量,全自动生化仪检测白蛋白和ALT水平,应用SPSS 19.0软件对数据进行统计学处理。结果:77例患者中,丙肝抗体阳性56例,阴性21例;HCV-RNA阳性71例,阴性6例。这2种方法检测出的结果差异有统计学意义(P0.05)。将45例HCV-RNA阳性的标本按拷贝数不同分成6组,检测各组白蛋白的变化,前4组随着HCV-RNA拷贝数的增加,白蛋白含量依次降低。57例HCV-RNA阳性的标本中,有22例ALT含量异常,并且随着HCV-RNA拷贝数的增加,ALT的异常率及平均值也在上升;经相关性分析,ALT平均值与HCV-RNA拷贝数成正相关(r=0.999,P=0.032)。结论:在丙型肝炎的诊断中,HCV-RNA拷贝数是重要的检测指标,同时应联合检测丙肝抗体、白蛋白和ALT。     

α2b-IFN诱导CD25表达和对PBMC内HCV RNA的阴转作用

目的探讨α2b-IFN对PBMC内CD25的诱导及对HCV-RNA、抗-HCV转阴效果。方法用BSA法检测慢性丙肝患者治疗前后CD25表达水平。以α2b-IFN(3 MU/d)治疗3个月为一疗程,共2个疗程,于治疗前后分别检测患者PBMC内HCV-RNA和血清HCV-RNA、抗-HCV。结果慢性丙肝患者α2b-IFN治疗后静息期、诱导期CD25表达水平分别为(3.44±0.77)%、(33.62±3.95)%,PBMC、血清内HCV-RNA和抗-HCV转阴率分别为42.31%(11/26)、57.69%(15/26)和65.38%(17/26),与对照相比差异有显著性(P<0.01~P<0.05)。结论α2b-IFN可诱导CD25表达,对PBMC内HCV-RNA具有肯定的治疗作用,其疗效优于常规治疗。     

核酸纯化柱提取核酸定量检测丙型肝炎病毒RNA的临床应用

目的:探究核酸纯化柱提取核酸定量检测血浆标本中丙型肝炎病毒核糖核酸(HCV-RNA)的临床应用效果。方法:将2013年8-11月期间我院600例抗-HCV阳性丙肝患者的样本,按随机字数表法分为研究组对照组各300例,分别采用核酸纯化柱和酚-氯仿提取法检测,采用实时荧光定量聚合酶链反应(FQ-PCR)技术定量检测两组HCV-RNA水平。结果:研究组检测阳性率为87.67%(263/300),显著高于和对照组的54.0%(162/300),差异有统计学意义(X2=82.296,P=0.000);两组HCV-RNA检测水平差异无统计学意义(u=1.721,P=0.067)。结论:核酸柱提法定量检测血浆HCV-RNA操作简单、快速,分离效率高,容易掌握,值得临床推广。     

类风湿因子分型(IgG、IgM、IgA)与丙型肝炎关系的初探

为了了解类风湿因子(RF)自身免疫抗体的分型(IgG、IgM、IgA)与丙型肝炎之间的关系,探讨其出现的规律及与肝功能损坏情况的相关性.采用定量双抗原夹心ELISA法进行检测.结果发现抗-HCV阴性、ALT＜40U的C组RF总阳性率为5%;抗-HCV阳性、ALT＜40U的A组为56%:抗-HCV阳性,ALT≥40U的B组为89%,C组与A、B组之间有显著性的差异P＜0.01).A组lgM-RF阳性率为4%;B组为56%;A组与B组之间有显著性差异(P＜0.05).因此抗-HCV阳性者血清中各型RF总阳性率比正常对照的C组有明显的增高;B组的IgM-RF比A组明显地增高.研究结果提示:IgM-RF的增高与丙肝患者肝功能损坏的发生与否存在密切的关系.     

慢性丙型肝炎患者治疗前后血清中IL-18、IFN-γ及IL-4的表达及意义

目的:研究慢性丙型肝炎患者治疗前后血清中白介素-18(IL-18)、干扰素-γ(IFN-γ)及白介素-4(IL-4)的表达情况,探讨IL-18、IFN-γ在及IL-4在慢性丙型肝炎发病中的作用及可能临床意义。方法:酶联免疫吸附法(ELISA)检测20例正常人,42例慢性丙型肝炎患者治疗前后血清中IL-18、IFN-γ在及IL-4水平。结果慢性丙型肝炎患者血清IL-18表达高于健康对照组(380.3±27.2pg/mlvs104.1±10.9pg/ml,P<0.05),血清IFN-γ表达也高于健康对照组(3.2±0.4IU/mlvs1.2±0.2IU/ml,P<0.05),而慢性丙型肝炎患者与健康对照组间IL-4表达无统计学差异(23.8±2.7pg/mlvs23.5±2.9pg/ml,P>0.05)。慢性丙型肝炎患者血清IL-18表达与谷丙转氨酶具有正相关性(r1=0.701,P<0.05);IFN-γ表达与谷丙转氨酶均具有正相关性(r2=0.629,P<0.05)。治疗前慢性丙型肝炎患者应对组血清中IL-18及IFN-γ表达高于无应答组(380.3±27.2pg/mlvs280.1±19.8pg/ml,P<...     

白介素-32在HBV感染者的变化及其临床意义

目的:探讨乙型肝炎病毒(HBV)感染者血清中白介素-32(IL-32)水平的变化及其临床意义。方法:对100例HBV感染者和30例健康者检测外周血清白细胞介素-32(IL-32)的含量,比较IL-32水平变化的规律,并分析其与临床疾病谱和HBV复制水平的关系。结果:(1)急性乙型肝炎、中度慢性乙型肝炎、重度慢性乙型肝炎患者血清IL-32水平与对照组有显著性差异(P<0.001);轻度慢性乙型肝炎患者血清IL-32水平高于健康对照组,但无统计学意义(P>0.05);急性乙型肝炎、中、重度慢性乙型肝炎患者血清IL-32水平均显著高于轻度慢性乙型肝炎组,且有统计学意义(P<0.001);急性乙型肝炎、重度慢性乙型肝炎患者血清IL-32水平均高于中度慢性乙型肝炎组,有统计学意义(P<0.001);急性乙型肝炎、重度慢性乙型肝炎组之间无明显差异(P>0.05)。(2)HBV感染者HBV-DNA阳性组IL-32水平较阴性组为高,但无差异;高、中、低病毒载量组之间血清IL-32水平也无差异(P>0.05)。(3)慢性乙型肝炎患者不同ALT水平组血清IL-32水平均高于对照组(P<0.05);且低、中、高ALT水平组之间与I...     

I b 型慢性丙肝患者血清IL-10、IL-12 水平 与HCV-RNA 载量关系的研究*

目的:研究I b型慢性丙肝患者血清IL-10、IL-12水平与HCV-RNA载量的关系,探讨难治性丙肝的免疫发病机制。方法:选取HCV-RNA阳性的I b型慢性丙肝患者48例,依据病毒载量将其分为三个组,低病毒载量组(1.0*103IU/mL1.0*106IU/mL)13例,另选HCV-RNA阴性的慢性HCV感染者15例,健康献血员15例作为对照组;应用ELISA法检测所有研究对象的血清IL-10、IL-12水平。结果:慢性丙肝患者外周血IL-10水平显著高于健康献血员(P=0.003),IL-12水平低于健康献血员(P=0.045);随着HCV-RNA载量的升高,HCV-RNA阳性的I b型慢性患者外周血IL-10水平逐渐升高,组间比较差异具有统计学意义(F=18.65,P=0.000);血清HCV-RNA载量与血清IL-10水平呈正相关(相关系数r=0.71,P=0.000),与血清IL-12水平无相关性(P=0.479)。结论:慢性HCV感染者与患者的细胞免疫均倾向于TH2型;I b型慢性丙肝患者外周血IL-10水平与HCV-RNA病毒载量呈正相关。     

RF、GPI、抗CCP抗体联合检测对类风湿关节炎的诊断意义

目的:研究类风湿因子(rheumatoid factor,RF)、葡萄糖-6-磷酸异构酶(glucose-6-phosphate isomerase,GPI)和抗环瓜氨酸肽(anti-cyclic citrullinated peptide,anti-CCP)抗体联合检测对类风湿关节炎(rheumatoid arthritis,RA)的诊断意义.方法:收集未经治疗的128例RA患者(RA组)、117例其他风湿病人(非RA组)、74例健康人(正常对照组)血清,采用酶联免疫吸附法(ELISA)检测GPI、抗CCP抗体,采用速率散射比浊法检测RF、C反应蛋白(CRP)、补体(C3、C4)、免疫球蛋白(Iga、IgG、IgM),采用魏氏法测红细胞沉降率(血沉,ESR).结果:(1)GPI、抗CCP抗体诊断RA的特异性分别为91.09%、94.14%,与RF(78.53%)比有显著差异(P<0.01),敏感性分别为75.0%、75.8%,与RF(80.47%)比无统计学差异(P>0.05).(2)GPI和RF、抗CCP抗体和RF、GPI和抗CCP抗体两两组合检测,两个指标均为阳性诊断RA的特异性分别为94.24%、95.81%、96.34%,与单独检测RF比有显著差异(P<0.01);两个指标任一阳性诊断RA的敏感性分别为85.16%、85.94%、87.50%,与单独检测RF比无统计学差异(P>0.05),但与单独检测GPI或抗CCP抗体比有统计学差异(P<0.05)计学.(3)三个指标联合检测均为阳性诊断RA的特异性高达98.43%,与单独检测RF、GPI或抗CCP抗体比有统计学差异(P=0.000,P=0.001,P=0.018),任一阳性诊断RA的敏感性为91.41%,与单独检测RF、GPI或抗CCP抗体比有统计学差异(P=0.012,P=0;000,P=0.001).(4)GPI阳性的RA患者关节炎部位数、CRP、ESR水平明显高于GPI阴性RA患者.且差异有统计学意义(P<0.05).结论:GPI、抗CCP抗体诊断RA比RF更具特异性,三者联合检测可显著提高诊断RA的特异性和敏感性.此外,GPI还可作为RA活动性指标.     

乙肝病毒载量与血清标志物及ALT相关性研究

探讨了乙型肝炎病毒核酸(HBV-DNA)水平与乙型肝炎免疫标志物(HBVM)和丙氨酸氨基转移酶(ALT)的关系。分别采用实时荧光定量聚合酶链反应(FQ-PCR),酶联免疫法和连续监测法检测了345例血清标本HBV-DNA含量,HBVM(HBsAg、HBsAb、HBeAg、HBeAb、抗HBcIgM)表达及ALT水平。HBsAg、HBeAg(和抗HBcIgM)阳性患者HBV DNA阳性率要明显高于HBsAg、HBeAb(和抗HBcIgM)阳性患者、仅HBsAg阳性患者及HBsAb、HBeAb阳性患者(P<0.01)。血清HBeAg阳性标本HBV-DNA阳性率为98.7%,明显高于HBeAg阴性标本的61.6%(P<0.01),并且血清HBeAg阳性标本HBV-DNA含量(log值,7.42±1.43)也明显高于HBeAg阴性标本(4.36±1.73)(P<0.01);在HBV-DNA含量小于107copy/mL的标本中,ALT与HBV-DNA含量呈正相关(P<0.01)。血清中HBV DNA含量与乙型肝炎免疫标志物以及肝细胞损伤三者之间存在密切的关系,在临床工作中应对血清HBVM、ALT和HBV-DNA含量联合检测,这样才能更准确地判断患者病情、预后及指导抗病毒药物的应用。     

艾滋病合并丙型肝炎的抗病毒治疗临床观察

目的:观察艾滋病合并丙型肝炎的抗病毒治疗效果并回顾总结艾滋病合并丙肝的临床研究进展。方法:选择艾滋病合并丙型肝炎的患者33例,采用HAART疗法及干扰素联合利巴韦林治疗的方法,治疗4周后观察以上患者谷丙转氨酶(ALT)、谷草转氨酶(AST)、谷氨酸脱氢酶(GLDH)、总胆红素(TBIL)、白蛋白(ALB)和HCV-RNA检查结果,同时总结艾滋病合并丙肝的临床研究进展。结果:治疗4周后,以上患者ALT、AST、GLDH、TBIL、ALB和HCV-RNA阳性患者显著改善,治疗前后比较差异具有(高度)统计学意义(P<0.05,P<0.01)。结论:在对艾滋病合并丙型肝炎充分了解的同时,积极采取抗病毒治疗措施,可延缓疾病的发展。     

Evaluation of an enzyme immunoassay for hepatitis C virus antibody detection using a recombinant protein derived from the core region of hepatitis C virus genome

This study was undertaken to evaluate an enzyme immunoassay (EIA) for hepatitis C virus antibody detection (anti-HCV), using just one antigen. Anti-HCV EIA was designed to detect anti-HCV IgG using on the solid-phase a recombinant C22 antigen localized at the N-terminal end of the core region of HCV genome, produced by BioMérieux. The serum samples diluted in phosphate buffer saline were added to wells coated with the C22, and incubated. After washings, the wells were loaded with conjugated anti-IgG, and read in a microtiter plate reader (492 nm). Serum samples of 145 patients were divided in two groups: a control group of 39 patients with non-C hepatitis (10 acute hepatitis A, 10 acute hepatitis B, 9 chronic hepatitis B, and 10 autoimmune hepatitis) and a study group consisting of 106 patients with chronic HCV hepatitis. In the study group all patients had anti-HCV detected by a commercially available EIA (Abbott), specific for HCV structural and nonstructural polypeptides, alanine aminotransferase elevation or positive serum HCV-RNA detected by nested-PCR. They also had a liver biopsy compatible with chronic hepatitis. The test was positive in 101 of the 106 (95%) sera from patients in the study group and negative in 38 of the 39 (97%) sera from those in the control group, showing an accuracy of 96%. According to these results, our EIA could be used to detect anti-HCV in the serum of patients infected with hepatitis C virus.     

Hepatitis C virus-RNA, immunoglobulin M anti-HCV and risk factors in haemodialysis patients

In order to evaluate hepatitis C virus-RNA (HCV-RNA), immunoglobulin M (IgM) anti-HCV and risk factors in haemodialysis patients, 180 subjects (45 HCV negative and 135 HCV positive) were studied. Sex, age, duration of dialysis, number of transfusions and ALT were also considered. HCV-RNA was determined by the Amplicor HCV test, and IgM anti-HCV by the Abbott HCV IgM EIA. These markers were present in 40% and 30.4% of anti-HCV positive subjects. The agreement between the two tests employed was 77%. The results showed a close association between HCV-RNA and IgM anti-HCV with abnormal ALT levels and between HCV-RNA and the number of transfusions. Both of these markers were different when correlated with age and time on dialysis, respectively. Therefore, IgM anti-HCV may also serve as a serological marker of HCV infection and a complementary marker of virus replication.     

乙型肝炎病人重叠感染丙型肝炎的评价

应用ＥＬＩＳＡ和ＰＣＲ法检测５０２例乙肝病人血清,４０１例ＨＢｓＡｇ阳性血清中,有１１４例（２８．４％）抗－ＨＣＶ和ＨＣＶＲＮＡ双项阳性,２５例（６．２％）ＨＣＶＲＮＡ单项阳性;２１例（５．２％）抗－ＨＣＶ单项阳性。将ＨＢｓＡｇ乙肝病人分成ＨＢＶＤＮＡ,ＨＢｅＡｇ阳性组和ＨＢＶＤＮＡ,ＨＢｅＡｇ阴性组。前者抗－ＨＣＶ阳性率为１１．６％～２０．５％,ＨＣＶＲＮＡ阳性率为１６．２％～２０．５％。后者抗－ＨＣＶ阳性率为２０．２％～５５．６％,ＨＣＶＲＮＡ阳性率为２３％～６０．３％。结果说明长期携带ＨＢＶ者和慢性乙肝病人均可重叠ＨＣＶ感染。ＨＢＶＤＮＡ阳性组抗－ＨＣＶ和ＨＣＶＲＮＡ阳性率明显高于ＨＢＶＤＮＡ阳性组     

Occult HBV infection and suppression of HCV replication in the early phase of combination therapy for chronic hepatitis C

BACKGROUND: Occult HBV infection in subjects with chronic hepatitis C is related to more severe disease outcome. It has been suggested that it might reduce sensitivity to antiviral treatment. AIMS: To assess in HBsAg negative subjects with chronic hepatitis C any effect of the presence of HBV genomes in the liver on the early kinetics of HCV-RNA under PEG-IFN plus ribavirin. PATIENTS AND METHODS: Twenty-two anti-HCV and HCV-RNA positive subjects, with biopsy-proven chronic hepatitis C (M/F 15/7; 50 +/- 8.6 years, 16 genotype 1b) were given PEG-IFN alpha 2b 1.0 microg qw plus ribavirin (800 to 1,200 mg daily according to body weight) for an intended 52 week period. Early virological response was assessed over the first 4 weeks of therapy by quantifying HCV-RNA. Occult HBV infection was assessed by testing for HBV-DNA in the liver before therapy. RESULTS: HBV genomes were found in the liver of 7 of 22 (31.4%) patients, unrelated to anti-HBc status. Kinetics of HCV-RNA during the first 4 weeks of antiviral treatment was unaffected by occult HBV infection, both in terms of absolute reduction of viral load and of number of cases with a reduction of > or = 2 log10 on treatment. CONCLUSIONS: Occult HBV infection does not affect the early phase of response to combination therapy. Further follow-up of patients into the maintenance phase of antiviral treatment and after stopping it will clarify if and when occult HBV has a role in reducing sustained virological response.     

丙型肝炎患者血浆和外周血单个核细胞同步检测HCV-RNA和HGV-RNA

To understand the incidence of hepatitis C and hepatitis G coinfection, and positive rate of HCV-RNA or HGV-RNA in plasma and PBMC, HCV-RNA and HGV-RNA in plasma and in peripheral blood mononuclear cells(PBMC) of the 40 patients were amplified with RT-PCR. There were 6 and 8 HGV-RNA positive cases in plasma and PBMC, respectively. And there were 5 HGV-RNA positive cases both in plasma and PBMC.At the same time, there were 5,6,3 both HCV-RNA and HGV-RNA positive cases in plasma, in PBMC and in both plasma and PBMC,respectively. It accounted for 13%,15% and 8%, respectively. The HCV-RNA and HGV-RNA positive incidence of PBMC was higher than that of plasma. The incidence of HCV-RNA and HGV-RNA coinfection was similar to European, American and Japanese. The synchronous detection has an important signficance in avoiding leaked diagnosis.     

Association between HLA class II antigens and hepatitis C virus infection

The aim was to confirm the influence of HLA Class II antigens on the progression of HCV infection and to assess the relationship between these antigens and histological damage, HCV viral load and HCV genotype. 143 patients were enrolled and divided into three groups. Group A included 34 anti-HCV positive, HCV-RNA negative patients with ALT persistently normal; group B included 39 patients with HCV-RNA positive and abnormal ALT level; group C included 70 normal subjects. Serological HCL typing was performed with lymphocytotoxicity test by Terasaky and McClelland, using lymphobeads HLC class II. The frequency of HLA DR11 (5) was significantly higher in the control group (52.9%) and in group A (64.7%), than in group B (28.2%). Allele HLA DR6 was demonstrated in a similar proportion (26%) among control group and group B, while HLA DR14 (6) was less frequent among controls (18% vs 1.4%). In group A the frequency of HLA DR14 (6) was 3% compared to group B, HLA DR17 (3) was prevalent (15.4%) in group B. Liver damage was associated with the detection of HLA DR14 (6) and HLD DR17 (3) antigens. Significantly lower levels of HCV-RNA were measured in subjects with HLA DR11 (5) than in these with either DR6 or DR17 (3). HLA class II antigens appear crucial for resolution or progression of HCV hepatitis. The punctual identification of these genetic factors may, therefore, prove to be useful in predicting disease evolution, in guiding the appropriate therapy for patients with poor prognosis, and in encouraging the development of now therapeutic strategies.     

Anti-hepatitic C virus antibodies hidden in circulating antibody/antigen aggregates in HCV-RNA positive patients

The aim of this study was to determine whether antibodies to HCV can be hidden in immunocomplex aggregates in anti-hepatitis C virus (HCV) negative, HCV-RNA positive patients and whether their presence could be related to HCV viral load or HCV genotype. Sera (23 in toto) from patients with elevated alanine aminotransferase (ALT) levels and negative for anti-HCV but positive for HCV-RNA and the immunocomplex aggregates (precipitate with PEG 6000 and glycine 1 M) were studied. The sera were treated using a rapid, simple new ELISA which disrupted the immunocomplex aggregates. Sera from ten patients were tested anti-HCV positive after immunocomplex disruption. No correlation with age, sex, ALT level, viral load or HCV genotype was observed. In some patients anti-HCV antibodies were hidden in circulating antibody/antigen complexes which could be dissociated with a simple, inexpensive and rapid protocol; therefore it can provide a valuable addition to the diagnosis of HCV infection and it may prevent some cases of post-transfusion hepatitis.     

IgM anti-hepatitis C virus in patients with chronic non-A, non-B hepatitis and their relationship to viral replication

Patients with hepatitis C virus (HCV) infection may have different patterns of antibody response to various structural and non-structural viral antigens. We have correlated the serological patterns to the clinical features of chronic infection and to viral replication in 68 HCV-Ab-positive patients with chronic liver disease at different stages (19 with cirrhosis-hepatocellular carcinoma, 38 with chronic active hepatitis and 11 with chronic persistent hepatitis). Serum samples from each patient were assayed for HCV-IgM by enzyme immunoassay and for HCV-RNA by the polymerase chain reaction using primer sets derived from the 5'-non-coding region. The prevalence of HCV-IgM was high (54 patients (79.4%)) and the study showed a good correlation between high values of anti-HCV-IgM and the presence of HCV-RNA in serum, since HCV-RNA was detected in 35 of the 54 IgM-positive patients (64.8%) and notably in 19 of the 20 subjects with high levels of specific IgM. Conversely, all the 35 sera containing HCV-RNA were also reactive for HCV-IgM, while none of the HCV-IgM-negative sera was HCV-RNA reactive. Positivity rates for both HCV-RNA and IgM anti-HCV were higher in the more advanced stages of disease; thus, the clinical pattern of HCV chronic hepatitis seems to be strictly related to the serological pattern and the presence of HCV-RNA.