IMPROVED QUANTITATIVE COMMERCIAL HIV ASSAY

THE DETECTION AND QUANTIFICATION OF INFECTIOUS HUMAN IMMUNODEFICIENCY VIRUS (HIV) WILL BE IMPORTANT FOR MONITORING BLOOD PRODUCTS, VIRUS LOAD IN PATIENTS, AND SCREENING ANTIVIRAL DRUGS. WE HAVE DEVELOPED A SPECIFIC AND SENSITIVE "INDICATOR CELL" BIO-ASSAY FOR THE DETECTION AND QUANTIFICATION OF INFECTIOUS HIV AND HIV-INFECTED CELLS. THE BIO-ASSAY CONSISTS OF A HUMAN CD4+ INDICATOR CELL LINE CONTAINING THE BETA-GALACTOSIDASE REPORTER GENE UNDER THE CONTROL OF HIV REGULATORY ELEMENTS. IN THE ABSENCE OF HIV, THE BETA-GAL GENE IS NOT EXPRESSED, HOWEVER, UPON EXPOSURE TO INFECTIOUS HIV, REPORTER GENE EXPRESSION IS ACTIVATED. THE SUP BETA-GAL INDICATOR CELL LINE EMPLOYED TO TITER INFECTIOUS HIV, HAS EXHIBITED 20-50 FOLD GREATER SENSITIVITY FOR HIV DETECTION COMPARED TO THE SYNCYTIA FORMATION ASSAY. THIS CELL LINE HAS ALSO BEEN USED TO DETECT HIV NEUTRALIZING ANTIBODY. THEY WISH TO OPTIMIZE THE IN VITRO ASSAY CONDITIONS, EXTEND THE CLINICAL COVERAGE OF THE ASSAY, AND CONSTRUCT ASSAYS USING OTHER REPORTER GENES TO ALLOW MORE RAPID QUANTITATION OF SIGNAL. THE OBJECTIVE IS TO DEVELOP A COMMERCIAL INDICATOR CELL ASSAY WHICH HAS BROAD CLINICAL COVERAGE AND IS HIGHLY REPRODUCIBLE AND SIMPLE PERMITTING ITS USE IN CLINICAL RESEARCH AND PATHOLOGY TEST LABORATORIES.