Rapid and sensitive test for detecting hepatitis A virus

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage

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536 2432, 536 2433, C12Q 170, C12P 1934, C07H 2104

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active

052906779

ABSTRACT:
Provided are methods for detecting hepatitis A virus (HAV) and HAV genotypes I and III by capturing whole hepatitis A virus with antibodies specific to hepatitis A virus, generating a cDNA copy of the RNA by reverse transcription in the presence of a negative strand primer having the sequence defined by SEQ ID NO:1, amplifying the cDNA by polymerase chain reaction with both the negative strand primer and a labelled or unlabelled positive strand primer having the sequence defined by SEQ ID NO: 2, and detecting the amplified cDNA by hybridization with either or both of the probes defined by SEQ ID NO:3 and SEQ ID NO:4 or by detection of label, wherein the presence of detectable hybridization or amplification indicates the presence of hepatitis A virus. Also provided are DNA primers and probes from the amino-terminal portion of the VPI region of the HAV genome that selectively hybridize with HAV, HAV genotype I and HAV genotype III and can amplify the amino-terminal portion of the VP1 region of HAV.

REFERENCES:
patent: 5075212 (1991-12-01), Rotbart
Cohen et al. (1987) Complete Nucleotide Sequence of Wild-Type Hepatitis A Virus: Comparison with Different Strains of Hepatitis A Virus and Other Picornaviruses, J. of Virology 61(1), 50-59.
Jansen et al. (1985) Combined Immunoaffinity cDNA-RNA Hybridization Assay for Detection of Hepatitis A Virus in Clinical Specimens J. Clinical Microbiology 22(6), 984-989.
Brown & Robertson (1990) Immunoselection of Clinical Specimens Containing Virus Followed by Polymersase Chain Reactor Amplification and Rapid Direct Sequencing Biotechniques 8(3), 262-264.
Robertson et al. (1991) Epidemiologic Patterns of Wild-Type Hepatitis A Virus Determined by Genetic Variation J. Infectious Diseases 163, 286-292.
Hayashi et al. (1989) Use of Labeled Primers in Polymerase Chain Reaction (LP-PCR) for a Rapid Detection of the Product Nucleic Acids Res. 17, 3605.
Brown et al., (1989) Characterization of a Simian Hepatitis A Virus (HAV): Antigenic & Genetic Comparison with Human HAV J. Virology 63(11), 4932-7.

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