36 results about "Hepatitis C virus genotype" patented technology

The invention relates to a kit for detecting the genotype and IL28-site polymorphism of a hepatitis C virus (HCV), in particular to a kit for detecting the genotype and IL28-site polymorphism of an HCV, which is prepared by using a nucleic acid reverse dot hybridization technology. The kit comprises an amplification reagent, a low-density chip using a nylon membrane as a carrier and a hybridization reagent, and can simultaneously detect the genotype of the HCV and the IL28B gene polymorphism closely related to the HCV treatment, thereby providing a more reliable basis for the individual hepatitis C treatment.

The invention relates to a kit for detecting a hepatitis c virus genotype, particularly relates to that the nucleic acid reverse dot blot hybridization technology is used to prepare a kit for hepatitis c virus genotype detection. The invention is used for rapidly and accurately distinguishing the hepatitis c virus genotype in a clinical blood sample.

The present inventors developed hepatitis C virus 1a/2a and 1b/2a intergenotypic recombinants in which the JFH1 structural genes (Core, E1 and E2), p7 and NS2 were replaced by the corresponding genes of the genotype Ia reference strain H77C or TN or the corresponding genes of the genotype Ib reference strain J4. Sequence analysis of recovered 1a/2a and 1b/2a recombinants from 2 serial passages and subsequent reverse genetic studies revealed adaptive mutations in e.g. p7, NS2 and/or NS3. In addition, the inventors demonstrate the possibility of using adaptive mutations identified for one HCV isolate in generating efficient cell culture systems for other isolates by transfer of mutations across isolates, subtypes or major genotypes. Furthermore neutralization studies showed that viruses of e.g. genotype 1 were efficiently neutralized by genotype Ia, 4a and 5a serum, an effect that could be utilized e.g. in vaccine development and immunological prophylaxis. The inventors in addition demonstrate the use of the developed systems for screening of antiviral substances in vitro and functional studies of the virus, e.g. identification of receptors required for HCV entry

The present inventors developed hepatitis C virus 6a/2a intergenotypic recombinants in which the JFH1 structural genes (Core, E1 and E2), p7 and the complete NS2 were replaced by the corresponding genes of the genotype 6a reference strain HK6a. Sequence analysis of recovered 6a/2a recombinants from 2 transfection experiments and subsequent reverse genetic studies revealed adaptive mutations in E1 and E2. Conclusion: The developed 6a/2a viruses provide a robust in vitro tool for research in HCV genotype 6, including vaccine studies and functional analyses.

Genotype 7a has been identified recently, thus not much is known about the biology of this new, major HCV genotype. The present inventors developed hepatitis C virus 7a/2a intergenotypic recombinants in which the JFH1 structural genes (Core, E1 and E2), p7 and the complete NS2 were replaced by the corresponding genes of the genotype 7a strain QC69 and characterized them in Huh7.5 cells. Sequence analysis of 7a/JFH1 recombinants recovered after viral passage in Huh7.5 cells following 4 independent transfection experiments revealed adaptive mutations in Core, E2, NS2, NS5A and NS5B. In reverse genetic studies the importance of these mutations for improved growth kinetics was shown. Adapted 7a/JFH1 viruses showed growth kinetics, infectivity and RNA titers comparable to a previously developed 3a/JFH1 reference virus. Conclusion: The developed 7a/JFH1 viruses provide a robust in vitro tool for research in HCV genotype 7, including vaccine studies and functional analyses.

The present inventors developed 5a/2a intergenotypic recombinants in which the JFH1 structural genes (Core, E1 and E2), p7 and all of or part of NS2 were replaced by the corresponding genes of the genotype 5a reference strain SA13. Compared to the J6/JFH control virus, after transfection of in vitro transcripts in Huh7.5 cells, production of infectious viruses was delayed. However, in subsequent viral passages efficient spread of infection and HCV RNA titers as high as for J6/JFH were obtained. Infectivity titers were at all time points analyzed comparable to J6/JFH control virus. Sequence analysis of recovered 5a/2a recombinants from 2 serial passages and subsequent reverse genetic studies revealed adaptive mutations in p7, NS2 and/or NS3. Infectivity of the 5a/2a viruses was CD81 and SR-BI dependant, and the recombinant viruses could be neutralized by chronic phase sera from patients infected with genotype 5a. Conclusion: The developed 5a/2a viruses provide a robust in vitro tool for research in HCV genotype 5, including vaccine studies and functional analyses of an increasingly important genotype in South Africa and Europe.

The invention relates to a HCV (hepatitis c virus) genotype detection kit. The kit extracts a sample nucleic acid by using a magnetic bead method; by using a real-time fluorescence quantitative PCR (polymerase chain reaction) technology, and taking a highly conserved area of a HCV genome as an amplification target, a specific primer and a TaqMan probe are designed, then HCV genes are subjected to rapid and accurate genotyping detection on a real-time fluorescence PCR instrument by PCR amplification, and an internal standard is added in a system, so that false negative results can be effectively prevented.

The invention provides a core antigen fragment for detecting hepatitis C virus and an application of the core antigen fragment. The antigen level of different HCV genotype serum samples is detected by using an Abbott Architect HCV quantitative core antigen kit, the sensitivity of different genotypes is compared and detected, a sample of specific genotype which influences the detection sensitivity is analyzed, a polymorphic site of the core antigen which influences the detection sensitivity of the kit is found, and the amino acid sequence of the core antigen is as shown in SEQ ID NO.1-2. The amino acid fragment of the polymorphic site of the core antigen can serve as a molecular marker of HCV core antigen detection and is used for detecting the genotype of hepatitis C virus 3b. Moreover, the existence of the molecular marker can indicate that the sensitivity of the conventional quantitative antigen detection kit is reduced when the kit is used for detecting a sample of the genotype of hepatitis C virus 3b, normalization and standardization of the core antigen detection can be facilitated, and the core antigen fragment can be used for preparing a kit for detecting the HCV 3b genotype.

The invention relates to a hepatitis c virus (HCV) genotype detection kit. The kit utilizes a paramagnetic particle method to extract sample nucleic acid, and by adopting a real-time fluorescence quantitative PCR (polymerase chain reaction) technology, taking a highly conserved area of an HCV genome as an amplification target, and designing a specificity primer and a TaqMan probe, typing detection of the HCV gene can be rapidly and accurately carried out on a real-time fluorescence PCR instrument through PCR amplification.

The present invention provides an antibody that specifically binds to envelope protein 2 of HCV of genotype 2a but does not immunologically react with envelope protein 2 of HCV of genotype 1a.

The invention discloses an expression vector PVX-6His-CTBt-Bt for producing a multi-epitope vaccine of hepatitis c virus. The multi-epitope vaccine 6His-CTBt-Bt of hepatitis c virus expressed by the vector comprises a histidine tag, an orally-taken immunologic adjuvant cholera toxin B subunit (CTB) and six B cellular antigen epitopes Bp of refractory hepatitis c virus genetype 1. To facilitate high-efficiency expression of the vaccine in tobacco, the 6His-CTBt-Bt gene optimized by an artificially synthesized codon is cloned to a high-efficiency plant virus expression vector potato X virus vector PVX201 to obtain the recombinant expression vector PVX-6His-CTBt-Bt of the multi-epitope vaccine 6His-CTBt-Bt of the hepatitis c virus; the expression vector is inoculated to the tobacco; the tobacco is regarded as a reactor to produce the anti-hepatitis c virus vaccine which aims to the refractory hepatitis c virus, is easy to purify, and can achieve a good orally-taken immune effect and resist against virus infection in vitro.

The present inventors developed hepatitis C virus 2b/2a intergenotypic recombinants in which the JFH1 structural genes (Core, E1 and E2), p7 and the complete NS2 were replaced by the corresponding genes of the genotype 2b reference strain J8. Sequence analysis of recovered 2b/2a recombinants from 2 transfection experiments revealed that 2b/2a was genetically stable. Conclusion: The developed 2b/2a viruses provide a robust in vitro tool for research in HCV genotype 2b, including vaccine studies and functional analyses.

This invention provides a nucleic acid comprising, in the following order, a 5' untranslated region comprising a particular nucleotide sequence of the genome of hepatitis C virus genotype 3a; a nucleotide sequence encoding a particular amino acid sequence of an NS3 protein, a nucleotide sequence encoding a particular amino acid sequence of an NS4A protein, a nucleotide sequence encoding a particular amino acid sequence of an NS4B protein, a nucleotide sequence encoding a particular amino acid sequence of an NS5A protein, a nucleotide sequence encoding a particular amino acid sequence of an NS5B protein of the hepatitis C virus genotype 3a; and a 3' untranslated region comprising a particular nucleotide sequence of a genome of hepatitis C virus genotype 3a.

The invention discloses an amplification primer, a sequencing primer, a kit and a method for detection of hepatitis C virus genotyping. The amplification primer has the base sequences shown as SEQ IDNo:1 and SEQ ID No:2, and the sequencing primer has the base sequence shown as SEQ ID No:3. The method for detection of hepatitis C virus genotyping provided by the invention adopts the originally created primers for amplification and sequencing, and can be used for detection of common genotypes 1b, 2a, 3a, 3b and 6a of human hepatitis C virus.

The invention provides a hepatitis C virus chimeric replicon and its construction method. The hepatitis C virus (HCV) chimeric replicon is a chimeric replicon of HCV genotype 2a and genotype 3a, and non-structural protein 5b, and is named as HCV GT2a/GT3aNS5b. The construction method includes: taking pRep-JFH-1 as the basis, and replacing its NS5B gene with GT3a NS5B gene, thus obtaining an NS5B chimeric replicon. Compared with the existing chimeric replicons taking GT1b as the skeleton, the hepatitis C virus chimeric replicon provided in the invention has high transient transfection efficiency, and the replication efficiency of the replicon does not need to be improved by constructing a stale cell line. Thus, the invention provides a rapid technical route for construction of more HCV clinical sample chimeric replicons in the future.

Methods for the rapid detection of the presence or absence of Hepatitis C Virus (HCV) in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers and probes targeting HCV and kits are provided that are designed for the detection of HCV.