Patent ID: 6623919
Filing Date: 2003-09-23
Classification: C12Q

Abstract:
A method for detecting Hepatitis C Virus (HCV) RNA or Human Immunodeficiency Virus (HIV) RNA in a biological sample, said method comprising:(A) performing a reverse transcription reaction using RNA derived from said sample as a template and at least one reverse transcription primer that will prime reverse transcription of DNA from HCV RNA and at least one reverse transcription primer that will prime reverse transcription of DNA from HIV RNA to produce reverse transcription products comprising (a) HCV-specific reverse transcription products, (b) HIV-specific reverse transcription products, or (c) a combination of (a) and (b); (B) amplifying said reverse-transcription products using one or more pairs of oligonucleotide primers specific for the 5â€² noncoding region of HCV and one or more pairs of oligonucleotide primers specific for HIV to produce amplification products comprising (a) HCV-specific amplification products, (b) HIV-specific amplification products, or (c) a combination of (a) and (b); wherein each of said pairs of oligonucleotide primers specific for HCV comprises: (i) forward primer 5â€²-GGGAGAGCCATAGTGGTCTGCGGAA-3â€² (C131F25) , and (ii) reverse primer 5â€²-CGGGGCACTCGCAAGCACCCTATCA-3â€² (C294R25) ; wherein each of the pairs of oligonucleotide primers specific for HIV-1 comprises a forward primer with the sequence: 5â€²-CTGCTTAAGCCTCAATAAAGCTTGCCTTGA-3â€² (JBLTR4) , and a reverse primer specific for HIV-1 selected from the group consisting of: (1) 5â€²-GGGTCTGAGGGATCTCTAGTTACC AGAGT-3â€² (JBLTR6) , and (2) 5â€²-TGTTCGGGCGCCACTGCTAGAGA-3â€²(JBLTR8) , wherein each of the pairs of oligonucleotide primers specific for HIV-2 comprises a forward primer with the sequence 5â€²-GGGAGGTTCTCTCCAGCACTAGCA-3â€² (2LTRe) , and a reverse primer specific for HIV-2 with the sequence 5â€²-GCGACTAGGAGAGATGGGAACACACA-3â€² (2LTR-R1) ; and (C) detecting said amplification products; wherein detection of HCV-specific amplification products indicates the presence of HCV RNA in said sample, detection of HIV-specific amplification products indicates the presence of HIV RNA in said sample, and the detection of HCV-specific amplification products and HIV-specific amplification products indicates the presence of HCV RNA and HIV RNA in said sample.