Simultaneous Detection of Multiplex-Amplified Human Immunodeficiency Virus Type 1 RNA, Hepatitis C Virus RNA, and Hepatitis B Virus DNA Using a Flow Cytometer Microsphere-Based Hybridization Assay

摘要: The feasibility of performing a multiplex assay for the detection human immunodeficiency virus type 1 (HIV-1) and hepatitis C (HCV) RNAs B (HBV) DNA is demonstrated. This based (i) on coamplification 142-bp fragment from gag region HIV-1 genome quantitation standard fragment, 244-bp 5' noncoding HCV genome, 104-bp pre-C gene regions HBV using three sets specific primers; (ii) capacity these four biotinylated PCR products to hybridize their oligonucleotide probe-coated microspheres; (iii) ability flow cytometer discriminate between distinct fluorescent-microsphere categories. Absence cross-hybridization unrelated probes generated by reverse transcription-PCR (RT-PCR) highly sensitive method allowed us assess unambiguously viral load infectious status 35 serologically well-established clinical samples 20 seronegative blood donor plasma tested. results indicate that RT-PCR microsphere-based hybridization assays, when combined, provide rapid, sensitive, major agents diseases in single sample.