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

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

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

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

Adult ; Asian Continental Ancestry Group ; DNA, Viral ; blood ; Female ; Genes, Viral ; genetics ; Hepatitis B virus ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Protein Isoforms ; genetics ; Viral Core Proteins ; genetics

OBJECTIVETo examine the antigenicity of hepatitis C virus (HCV) F protein and investigate serum prevalence of anti-F in HCV-infected patients.

METHODSEleven pairs of overlapping primers were used to synthesize the full-length HCV f gene, from which the truncated HCV f65 gene fragment was amplified by PCR. HCV f65 gene was then cloned into pET32a(+), and transformed into E. coli strain Plyss (DE3). This recombinant E.coli was induced by IPTG for the production of HCV F65 protein. The expressed HCV F65 protein, purified by Ni-NTA agarose, was further used in ELISA to detect serum anti-F, and to immunize rabbits for making polyclonal anti-F. The rabbit polyclonal anti-F was purified by Staphylococcus aureus protein A agarose.

RESULTSAfter recombinant pET32a(+)-f65 was constructed successfully, HCV F65 protein was expressed and purified. The purified HCV F65 protein was used as a capture antigen in ELISA to detect serum anti-F in HCV infected patients (n = 30). The result showed that the mean A450 value and the positive rate of serum anti-F were 0.125+/-0.061 and 63.3%, respectively. The rabbit-derived polyclonal anti-F reacted specifically with HCV F65 protein, of which the titer was 1:30,000.

CONCLUSIONOur expressed HCV F65 protein is of antigenicity, and can be used to determine serum anti-F. Anti-F IgG does exist in the sera of the HCV-infected patients. Moreover, the rabbit-derived polyclonal anti-F can be used to detect HCV F protein.

Animals ; Antibodies, Viral ; blood ; Hepacivirus ; genetics ; immunology ; Hepatitis C ; blood ; epidemiology ; immunology ; Hepatitis C Antigens ; blood ; immunology ; Humans ; Prevalence ; Rabbits ; Viral Core Proteins ; blood ; immunology ; Viral Envelope Proteins ; immunology

BACKGROUNDTo observe the features of serum specific IgA, IgG, IgM antibodies in the acute phase of hemorrhagic fever renal syndrome (HFRS).

METHODSThe nucleocapsid (NP) protein and glycoproteins (GP) of Hantavirus were expressed by recombinant baculovirus, and used as ELISA antigens to test 61 serial sera of 14 acute phase HFRS patients.

RESULTSSeoul like virus RNA were detected from 11 of 14 patients. An early and strong IgA, IgG and IgM antibody response to recombinant NP (rNP) was observed in almost all HFRS cases. The titers of antibody to rNP was apparently higher than that to Rgp. In the early stage, titer of IgG antibody elevated most drastically among all the three classes of antibodies to rNP, followed by IgM and IgA antibody responses. The elevation trend of IgM and IgA antibodies to rNP stayed nearly at the same level, but the IgA titers to rNP were apparently higher than that of IgM. Among the antibodies to rGP, IgA changed distinctly greater than IgG. The elevation trend of IgM could be found during first week after the onset, and the titers dropped gradually after the second week. IgM antibodies of one case who was viral RNA positive were not detected at early stage, but IgA titers were high. The only severe case of the 14 patients kept the lower IgA, IgG and IgM during the whole acute phase.

CONCLUSIONHFRS patients kept an early and strong humoral response to NP and GPs in acute phase of HFRS.IgA could be used together with IgM to improve the diagnostic accuracy.

Acute Disease ; Adult ; Antibodies, Viral ; blood ; Capsid Proteins ; genetics ; immunology ; Female ; Hantavirus ; genetics ; immunology ; Hemorrhagic Fever with Renal Syndrome ; immunology ; virology ; Humans ; Immunoglobulin Isotypes ; blood ; Male ; Viral Core Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology

This study was purposed to investigate the feasibility to screen donor with HCV infection by means of HCV-cAg ELISA. The first and repeat assays were performed for detection of serum anti-HCV in 8677 donor's serum specimens from January 2003 to December 2005. All serum anti-HCV specimens with positive anti-HCV from first and repeat assays were finally identified by using HCV-cAg ELISA and HCV RT-PCR methods. The results showed that only 5 serum specimens were positive anti-HCV by HCV-cAg ELISA identification in 29 specimens including 15 specimens with positive ant-HCV in first assays and 14 specimens with positive anti-HCV in repeat assays, the positive rate detected by HCV cAg ELISA was 17.24%. 5 serum specimens were positive anti-HCV by HCV RT-PCR detection also in 29 specimens mentioned above, the positive rate detected by HCV RT-PCR was 17.24% too. It is concluded that sensitivity of HCVcAg ELISA is similar to HCV RT-PCR and may be useful for the early diagnosis of hepatitic C or used as a reliable method to screen donor with HCV infection in blood transfusion medicine.
Blood Donors ; Blood Transfusion ; Enzyme-Linked Immunosorbent Assay ; methods ; Hepacivirus ; isolation & purification ; Hepatitis C Antigens ; blood ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Core Proteins ; blood

OBJECTIVETo investigate the variations of HCV core and E2 region epitopes during transfusion of activated immune cells.

METHODSFour patients receiving transfusion of activated immune cells were under continuously observation. HCV titers were measured by quantitive PCR. HCV core and E2 regions were cloned and sequences were analyzed by computer software.

RESULTSDuring the follow-up the serum ALT levels and the HCV virus titers fluctuated greatly in each of these persons. After transfusion, no significant variations were observed in HCV core and E2 coding regions.

CONCLUSIONSThe alteration of host immune attacks could induce the fluctuations of HCV load without any mutations in the currently observed coding genes of the epitopes.

Adult ; Aged ; Alanine Transaminase ; blood ; Epitopes ; genetics ; Female ; Genetic Variation ; Hepacivirus ; genetics ; Hepatitis C, Chronic ; blood ; therapy ; virology ; Humans ; Immunotherapy ; Male ; Middle Aged ; Viral Core Proteins ; genetics ; Viral Envelope Proteins ; genetics ; Viral Load

OBJECTIVETo investigate the genetic variation and typing of hepatitis B virus (HBV) in patients with chronic hepatitis B in relation to HBeAg status.

METHODSFluorescence quantitative polymerase chain reaction (PCR) was employed to detect serum HBV DNA in patients with chronic hepatitis B negative for HBeAg. Real-time fluorescent PCR and PCR-reverse dot blot hybridization were used to detect HBV genotypes and mutations, respectively.

RESULTSOf the 389 patients, 214 (55.01%) were positive and 175 (44.99%) were negative for HBV DNA; 102 (26.22%) had a HBV DNA copy number of 1×10(3), and 41 (10.54%) had a copy number of 1×10(4) (Χ(2)=226.6729, P<0.001). Of the 21 patients with a HBV DNA load of 1×10(5), 15 (71.43%) were found to have precore mutations, and 11 (52.38%) had basic core promoter (BCP) mutations; a higher HBV-DNA load was associated with an increased incidence of HBV mutations. In the 214 patients positive for HBV DNA, HBV genotypes A, B, C, D and the mixed type were found in 6 (2.80%), 84 (39.25%), 106 (49.53%), and 7 (3.27%), and 11 (5.14%) patients, who showed precore mutation rates of 16.67% (1 case), 36.90% (31 cases), 44.34% (47 cases), 0, and 0, and BCP mutation rates of 0, 19.05% ( 16 cases), 26.42% (28 cases), 0, and 0, respectively, demonstrating significant differences in HBV mutations between the genotype groups (P<0.001).

CONCLUSIONHBeAg-negative and HBV DNA-positive patients with chronic hepatitis B have a relatively low HBV replication level, and HBV DNA load is associated with HBV mutations. The B and C genotypes are more likely to have HBV mutations in HBeAg-negative patients.

DNA, Viral ; blood ; Female ; Genotype ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Male ; Mutation ; Promoter Regions, Genetic ; Viral Core Proteins ; genetics ; Viral Load