A RAPID AND SENSITIVE TEST FOR DETECTING HEPATITIS A VIRUS

摘要

A rapid and sensitive test for detecting Hepatitis A virus comprises a method of detecting the presence of HAV in a sample, comprising the steps of: (a) isolating RNA from a sample suspected of carrying HAV; (b) obtaining cDNA complementary to the isolated RNA of step (a); (c) amplifying the cDNA of step (b) by PCR utilizing one of a set of primers having the nucleotide sequences 5' GGAAATGTCTCAGGTACTTTCTTTG 3' and 5' GTTTTGCTCCTCTTTACATGCTATG 3' for amplification of the VP1 amino terminal genome region, or 5' CAACGGCCAGAGCCTAGGGCAAGGGG 3' and 5' CCCTCTTGGAAGTCCATGGTGAGGGGA 3' for the 5' non coding region; (d) detecting the amplified cDNA product by hybridization of a labeled internally located probe (VP1 (+) sense probe), nucleotides 22322252, 5' TCAACAACAGTTTCTACAGA 3'; VPI (-) sense probe, nucleotide 2283-2301, TAAATCTTTCATGGTTGT 3' or 5' non-coding region probe, nucleotides 161-187, 5' AGGTTCAGGGTTCTTAAATCTGTTC 3'; (e) alternatively, amplifying the cDNA described in step (c) by PCR in which one of the primers has been previously radiolabeled at the 5' end using polynucleotide kinase, followed by autoradiographic identification of the labeled product after agarose or acrylamide gel electrophoresis.