OBJECTIVE: To evaluate the clinical significance of serum hepatitis C virus (HCV) core antigen detected by enzyme linked immunosorbent assay (ELISA). METHODS: The serum HCV core antigen, which was taken from 149 patients with chronic hepatitis C, 20 patients of chronic hepatitis B and 20 health volunteers, was detected by ELISA. Meanwhile, the serum HCV RNA was detected by RT-PCR, and anti-HCV was detected by ELISA. RESULTS: The qualitative HCV core antigen in the serum, which was take from 20 patients of chronic hepatitis B and 20 health volunteers, was negative.The positive percentage of HCV core antigen was 49.66% in the 149 sera of patients with chronic hepatitis C. The coincidence of detective results of HCV RNA and HCV core antigen was 54.36%, without significant difference (P>0.05). The positive percentage of HCV RNA and HCV core antigen in the 149 anti-HCV antibody positive sera samples were 55.03% (82/149) and 49.66% (74/149), respectively, and there was no significant difference (P>0.05). CONCLUSION The qualitative HCV core antigen detected by ELISA has a high specificity. The positive percentage of HCV core antigen in the serum of patients with chronic hepatitis C is 49.66%. HCV core antigen is related to HCV RNA. HCV core antigen may be a useful serum marker which could show HCV viraemia like HCV RNA.
Hepatitis C Antigens ; blood ; Hepatitis C, Chronic ; blood ; Humans ; RNA, Viral ; blood ; Viral Core Proteins ; blood

OBJECTIVETo learn the prevalence of infection of human and animals severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) in Yantai, Shandong province, and to analyze the pathogenic features of SFTSV as well as its relationship between human and animal hosts.

METHODSFrom April to November in 2011, 3 576 serum samples were collected from domesticated animals, including sheep, cattle, pigs, dogs, chickens, in Laizhou and Penglai areas where fever with thrombocytopenia syndrome frequently occurred among local residents. Total SFTSV antibodies and virus-specific nucleic acids of the serum were tested by ELISA and Real time RT-PCR, respectively. SFTSV infection on each animal was observed in different months. 2 590 human serum samples were also collected in Laizhou and Penglai areas, with IgG antibodies tested by ELISA. Virus was isolated with Vero cells from the serum which SFTSV viral nucleic acids were positive. S fragments were amplified by RT-PCR and sequenced, with homology analysis conducted on these sequences.

RESULTSThe overall positive rate of serum samples from animals on the total SFTSV antibodies was 40.24% (1 439/3 576) while the positive rate for specific nucleic acids was 4.56% (163/3 576). The positive rates for SFTSV antibodies were 62.78%, 52.97%, 45.56%, 28.73%, 1.45% and the positive rates for specific nucleic acids were 5.72%, 4.63%, 3.02%, 5.25% and 3.73%, in sheep, cattle, chickens, dogs, pigs, respectively. The antigens/antibodies for SFTSV in animals changed seasonally. The overall positive rate for SFTSV IgG antibody from 2 590 human samples was 5.41%. Thirteen virus strains were isolated from these serum samples (10 strains from human and 3 strains from animals). The nucleotide homology of 13S fragments' sequences ranged from 95.23% to 100.00% and the nucleotide homology with the isolates from other provinces were between 94.72% and 99.13%. The homology was considered to be high.

CONCLUSIONHigh prevalence of SFTSV infections occurred both in human and domestic animals in Yantai city. The nucleotide sequences of SFTSV were highly homologous among human and domestic animals. The findings suggested that domesticated animals might serve as SFTSV proliferation and the hosts for transmission thus should be attached great importance.

Animals ; Antibodies, Viral ; blood ; Bunyaviridae Infections ; epidemiology ; China ; epidemiology ; Humans ; Prevalence ; RNA, Viral ; blood ; Sequence Analysis, RNA

China ; DNA, Viral ; blood ; Hepacivirus ; classification ; Hepatitis C ; diagnosis ; etiology ; therapy ; Humans ; RNA, Viral ; analysis

BACKGROUNDTo study the significance of the expression of HCV core protein in PBMC of patients with chronic hepatitis C and to evaluate the relationship between HCV core protein expression and clinical states.

METHODSIdentification of HCV protein antigen (Ag) in PBMC of 66 hepatitis C patients by immunohistochemical method and clinical status of the patients with HCV protein positive expression were investigated. In 27 out off 66 patients the HCV RNA and HCV Ag in PBMC were detected with RT-PCR and immunohistochemical method.

RESULTSThe HCV Ag (core+NS3) was identified in PBMC in 51 out of 66 patients (77.27%). It was also demonstrated that HCV core protein in nucleus showed strong expression and NS3 protein in cytoplasm showed weak expression. The expression of core protein in nucleus of PBMC were 35.29% in advanced chronic hepatitis patients, which was significantly higher than that from moderate cases (5.88%).

CONCLUSIONSThis study suggested that the expression of PMBC-HCV core protein may be related to the clinical state of the patients. The nuclear expression of HCV core protein in PBMC of patients with hepatitis C may be related to the persistence and activity of the chronic hepatitis C virus and play an important role in the pathogenesis of cirrhrosis and hepatocellular carcinoma.

Hepatitis C Antigens ; blood ; Hepatitis C, Chronic ; virology ; Humans ; Immunohistochemistry ; Leukocytes, Mononuclear ; metabolism ; RNA, Viral ; blood ; Viral Core Proteins ; blood

BACKGROUNDHepatitis C virus (HCV) core antigen assays have been produced to exclude infectious donations collected during the preseroconversion window phase (PWP). For the same purpose, we evaluated the specificity and sensitivity of a novel hepatitis C virus NS3 antigen detection immunoassay and the application of this assay in clinical diagnosis.

METHODSSamples from 77 healthy subjects, 173 anti-HCV positive patients and 3708 hepatitis patients other than HCV positive were tested with the HCV NS3 antigen assay. Some HCV NS3 antigen positive samples were further validated with HCV-RNA, neutralization and immunodot assays. Twenty-five sequential samples from 11 HCV NS3 antigen positive patients were subjected to kinetic study.

RESULTSOnly 48 (1.3%) of 3708 anti-HCV negative samples were positive for HCV NS3 antigen. Among them, 44 of 3030 samples from patients only infected with HBV were HCV NS3 antigen positive, 4 of the 445 samples from patients infected with other type hepatitis were HCV NS3 antigen positive. In addition, 42 (24.3%) of 173 anti-HCV positive samples were HCV NS3 antigen positive and all 77 samples from healthy subjects were negative to HCV NS3 antigen assay. Of the 15 HCV NS3 antigen positive samples, 9 (60%) were HCV-RNA positive. The neutralization and positive percentage of immunodot assay for 23 HCV NS3 antigen positive sera were 87.0% (20/23) and 69.6% (16/23) respectively. Of the 25 sequential samples from 11 HCV NS3 antigen positive patients, there was a negative correlation between the OD values and the duration of test (r = -0.989, P < 0.05), and there were correlations among their HCV NS3 antigen, HCV-RNA and anti-HCV titres. The anti-HCV antibodies of two sera were detected while their OD values of HCV NS3 antigen decreased gradually.

CONCLUSIONSThe HCV NS3 antigen detection assay showed perfect specificity and high sensitivity. Thus, it would be useful and economical as a routine test in laboratories for early diagnosis of HCV infection and prevention.

Alanine Transaminase ; blood ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; RNA, Viral ; blood ; Sensitivity and Specificity ; Viral Nonstructural Proteins ; blood

Hepacivirus ; classification ; genetics ; Hepatitis C ; diagnosis ; Hepatitis C Antibodies ; blood ; Humans ; RNA, Viral ; blood ; Viral Core Proteins ; blood

OBJECTIVETo investigate the value of detection of hepatitis C virus core antigen (HCV-cAg) for screening blood donor by using the internal reagent enzyme immunoassay (EIA) and anti-HCV antibody.

METHODSThe first and repeat assays were performed for detection of serum anti-HCV and HCV-cAg ELISA in 3972 donor's serum specimens from August to October of 2004. Twenty-five donors positive for anti-HCV were tested with HCV-cAg EIA kits and the results were compared with the results of HCV RNA determination with RT-PCR method.

RESULTSIn 3972 donor's serum samples, only 1 serum specimen was positive for HCV RNA identification among 10 specimens which were positive for anti-HCV in first assays, and only 1 serum specimens was positive for HCV RNA identification among 12 specimens positive for anti-HCV in repeat assays, only 2 serum specimens were positive HCV RNA identification in 3 specimens which were positive for HCV-cAg assays.

CONCLUSIONThe sensitivity of HCV-cAg ELISA is similar to HCV RT-PCR, but it is much cheaper. Therefore, HCV-cAg ELISA and anti-HCV may be used together to screen blood donor.

Blood Donors ; Enzyme-Linked Immunosorbent Assay ; Hepatitis C Antibodies ; blood ; Hepatitis C Antigens ; blood ; Humans ; RNA, Viral ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Core Proteins ; blood

No abstract available.
Hepatitis C/*diagnosis ; Hepatitis C Antibodies/*blood ; Humans ; Immunoenzyme Techniques ; RNA, Viral/*blood ; Viremia/*diagnosis

OBJECTIVETo detect humoral immune response against different function regions of hepatitis C virus (HCV) in chronic patients, and further to investigate the correlativity between anti-HCV antibody titers and HCV RNA concentration.

METHODSUsing recombinant dominate epitope antigens, e.g. HCV Core, NS3, NS4, NS5 and chimeric HVR1, a set of ELISA test reagents was formulated. Then, titers of antibodies against HCV different regions and the RNA concentration of HCV in chronic patient sera were detected by ELISA and quantitative RT-PCR technique, respectively.

RESULTSGreat differences have been noted in antibody titers and positive rate of different HCV function regions in chronic patients. Antibodies against HCV Core and HVR1 have the highest positive rate, then NS3, NS4, and NS5 in sequence.

CONCLUSIONThe titer of antibodies against different regions of HCV in chronic patients has good correlation with HCV RNA concentration.

Hepatitis C Antibodies ; blood ; Hepatitis C, Chronic ; immunology ; virology ; Humans ; RNA, Viral ; blood

OBJECTIVETo find out the mechanism of drug resistance by detecting the mutations of HIV RNA in patients who failed in the anti-HIV therapy, to direct the clinical use of anti-HIV drugs and to complement the existing drug resistant database.

METHODSHIV RNA and DNA were extracted from the plasma of 10 HIV-infected patients who developed drug resistance in the Clinic of AIDS, Ruhr University, Bochum, Germany. Then HIV-RNA was amplified in the reverse transcriptase (RT) and protease regions by polymerase chain reaction (PCR). After purified, the PCR products was sequenced. The acquired sequences were compared with the international standard strain HXB2CG and the resistant database of Stanford University.

RESULTSSome mutations were found to cause the corresponding resistance to certain drugs and were consistent with the clinical results. Some mutations existed in some patients, such as V179I in RT and K20T, K20I in protease, which hadn't been reported in the resistant database of Stanford University yet.

CONCLUSIONPatients who fail in HAART have different mutations in RT and protease regions. Mutations such as V179I in RT and K20T, K20I etc in protease may be related to drug resistance.

Adult ; Antiretroviral Therapy, Highly Active ; Drug Resistance, Viral ; HIV Infections ; drug therapy ; virology ; Humans ; RNA, Viral ; blood ; Treatment Failure