Adult ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; Humans ; Male ; Turkey

OBJECTIVETo investigate the relationship of hepatitis C virus (HCV) serotype with genotype.

METHODSThe serotypes of HCV in the serum of 104 patients with chronic hepatitis C from 14 cities in China for which HCV genotypes were available, were determined by ELISA using Murex HCV Serotyping 1-6 Assay.

RESULTSThe serotypes of 86 (82.69 percent) of the 104 serum specimens were determined, and HCV serotypes were determined for 91 strains. Overall the concordance between hepatitis C virus serotype and genotype was 62.1 percent, and the concordance of serotype, with genotypes 1, 2 and 3 were 69.4 percent, 51.2 percent and 70.0 percent, respectively. The false-negative rate and concordance of genotype 2b was lower (54.5 percent).

CONCLUSIONThe specificity of HCV serotyping was affected by HCV strains' genotype and sometimes HCV serotype was not in concordance with genotype.

Genotype ; Hepacivirus ; classification ; Hepatitis C, Chronic ; virology ; Humans ; Serotyping

OBJECTIVETo investigate the effect of hepatitis C virus (HCV) strain JFH1 on expression of the human gene, growth arrest and DNA damage-inducible gene 45 alpha (GADD45a), in infected hepatoma cells.

METHODSHCV JFH1 RNA-containing supernatants were used to infect the human hepatoma cell line, Huh7.5.1; infection was confirmed by Western blot detection of the HCV-encoded non-structural 5A (NS5A) protein and core protein. Infection-induced changes in GADD45a mRNA and protein expressions were measured by real time PCR using SYBR Green and Western blotting, respectively. Significance of differences between the levels detected in JFH1-infected or uninfected Huh7.5.1 cells was analyzed by single factor analysis of variance testing.

RESULTSThe HCV infection system was successfully established, as evidenced by expression of NS5A protein and core protein. The GADD45a mRNA and protein levels were significantly down-regulated in JFH1-infected Huh7.5.1 cells, by 0.57+/-0.09 and 0.28+/-0.03, respectively, as compared to levels in uninfected Huh7.5.1 cells (F values were 75.407 and 560.04, respectively; P less than 0.01).

CONCLUSIONHCV inhibits the mRNA transcription and protein expression of host GADD45a, which may contribute to the pathogenesis of hepatocellular carcinoma caused by HCV infection.

Cell Line, Tumor ; DNA Damage ; Hepacivirus ; classification ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Transcription, Genetic

BACKGROUND: There are several serological subtypes (serotypes) of hepatitis C virus (HCV) which can be detected by a serological enzyme immunoassay (EIA) method which detects the HCV subtype-specific antibodies against the NS4 region of the virus. We determined the HCV serotypes chronic HCV infected patients in terms of clinical applications. METHODS: Subtypes based on Simmonds' classification were detected in chronic HCV patients serologically and viral loads quantitated with branched DNA (bDNA) assay. The EIA was compared with multiplex PCR method. RESULTS: The serotype 1 (Simmonds' classification, 1a, 1b, 1c), the most prevalent form in Korea followed by serotype 2 (Simmonds' classification, 2a, 2b, 2c). The distribution of serotypes and HCV viral loads were not different according to disease severity. The patients infected with serotype 1 had higher viremic level (serotype 1, 7.4+/-15.5 MEq/mL; serotype 2, 1.1+/-2.2 MEq/mL, P=0.017). The serotype matched with the genotype in 85.7% (18/21) of cases. CONCLUSIONS: HCV serotype 1 is the most prevalent form in Korea and seemed to be able to more actively replicate than serotype 2. Both multiplex PCR and EIA can be used for detecting HCV subtypes and mutually interpretable.
Antibodies ; Classification ; DNA ; Genotype ; Hepacivirus ; Humans ; Immunoenzyme Techniques ; Korea ; Multiplex Polymerase Chain Reaction ; RNA* ; Viral Load

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

OBJECTIVEIn order to fully understand hepatitis c virus (HCV) genotype 3b, 1a, 2b and 6a infection in China, We built HCV 5'-noncoding region (5'-NCR) of different genotypes and subtypes.

METHODSThe classification HCV into variable genotypes (subtypes) was carried on by programs A, B and C A. Using a combination of three restriction endonuclease BHH' (BsrB I, Hae II, Hinf I) digestions at the same time. The distinct genotypes were classified into 5 groups: genotype 1 (1a, 1b), 6a, 2 (2a, 2b), genotype 3 (3a, 3b), genotype4 (4a). B. With regard to genotype 1, we could distinguish subtype 1a from 1b using BstU I digestion. C. Using restriction endonuclease Hae III, genotype 2a, 2b, 3b, 4a, 6a are differentiated respectively.

RESULTS(1) HCV genotype 1a, 1b, 2a, 2b, 3a, 3b, 4a, 6a are fully discriminated by comparison with the genotypes regular samples. (2) Of the 93 patients, HCV genotype distribution in China was 66.67% for 1b, 18.28% for 2a, 3.23% for 1b/2b, 3b, 2b respectively. 2.15% for 2a/2b, 1b/2a respectively. 1.08% for 1a.

CONCLUSIONThis research indicated that adoption of HCV 5'-NCR A B C restriction endonuclease digestions techniques, might be sensitive and efficient to detect HCV and discriminate HCV genotype (subtypes) 1a to 6a.

5' Untranslated Regions ; chemistry ; DNA Restriction Enzymes ; Genotype ; Hepacivirus ; classification ; genetics ; RNA, Viral ; analysis

CD4-Positive T-Lymphocytes ; HIV ; HIV Infections ; virology ; Hepacivirus ; classification ; Hepatitis C ; virology ; Humans ; Liver Transplantation ; Postoperative Period ; Superinfection ; virology

To investigate risk factors for HCV infection according to the genotype, we stud-ied 178 patients positive for HCV-PCR and 226 controls that were negative for the anti-HCV antibody. One hundred and twenty five controls (community con-trol) were recruited from spouses of HCV-PCR-positive patients and the other 101 from hospital visitors (hospital control). HCV genotyping was performed by PCR, and epidemiological data were obtained from all participants. The distribu-tion of HCV genotypes was as follows - 1a (0.6%), 1b (39.9%), 2a (38.2%), 2b (0%), 3 (1.1%), and unclassified (20.2%). By multivariate analysis, blood transfu-sion (OR 2.90) and endoscopy (OR 2.80) were found to be risk factors for HCV genotype 1b versus the community control. Similarly, blood transfusion (OR 3.17) was found to be risk factors for HCV genotype 1b versus the hospital control. Blood transfusion (OR 2.75) and endoscopy (OR 3.57) were risk factors for HCV genotype 2a versus the community control, and blood transfusion (OR 4.55) and endoscopy (OR 2.16) were those versus the hospital control. Our results suggest that the risk factors for HCV infection are similar among the different genotypes. Blood transfusion and endoscopy were found to be associated with HCV infection.
Female ; Genotype ; Hepacivirus/classification/genetics ; Hepatitis C/*virology ; Humans ; Korea ; Male ; Middle Aged ; Multivariate Analysis ; Risk Factors

OBJECTIVEUsing polymerase chain reaction-reverse blot dot (PCR-RDB) technique to establish a new method for hepatitis C virus (HCV) genotyping and to study the distribution of HCV genotypes in Foshan area.

METHODSHCV primers and probes were designed in 5'-untranslated region (nt-1-nt-299) of HCV. HCV RNA in serum was isolated and purified, and its cDNA was obtained by reversed transcription. Nested PCR using biotin-labelled primers, was done. PCR products were hybridized with immobilized specific probes (genotype 1a to 3b) on Biodyne C membrane to genotype HCV by color development while adding POD and TMB. A certain judgment could be made according to the position of color reaction. The reliability of this new method was verified by sequencing. HCV RNA levels in serum were determined by real time fluorescent quantitative (FQ)-PCR. 60 FQ-PCR-positive HCV sera from Foshan area were genotyped using this assay.

RESULTSAll 60 sera could be successfully genotyped by PCR-RBD. 50 (83.3%) cases were found to be genotype 1b, 2 (3.3%) as genotype 1a and 2 (3.3%) as genotype 2a while 5 (8.0%) to be mixture of genotype 1a and 1b, and 1 (1.7%) to be mixture of genotypes 1b and 2a. No genotypes 2b, 3a and 3b were found. The results of PCR-RDB genotyping methods coincided with sequence analysis.

CONCLUSIONNewly established HCV genotyping system was proved to be sensitive, specific, precise and economic, thus suitable for clinical and epidemiologic studies. The results of HCV genotyping showed that genotype 1b was the predominant genotype in Foshan area.

Genotype ; Hepacivirus ; classification ; genetics ; Hepatitis C ; virology ; Humans ; Immunoblotting ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo determine the distribution of HCV genotypes among volunteer blood donors in Guangzhou.

METHODSSix-nine HCV RNA-positive samples were collected from volunteer blood donors in Guangzhou. NS5B fragments of HCV were amplified followed by DNA sequencing and phylogenetic analysis.

RESULTSHCV genotypes were determined for 67 samples. Among them, the subtypes 1b, 2a, 3a, 3b, 6a and 6n were detected at the frequencies of 37.31%, 4.48%, 7.46%, 4.48%, 44.78% and 1.49%, respectively.

CONCLUSIONHCV 1b and 6a are the most predominant two subtypes among volunteer blood donors in Guangzhou.

Blood Donors ; China ; Genotype ; Hepacivirus ; classification ; genetics ; isolation & purification ; Humans ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, DNA