Relative quantitation of HIV-1 proviral DNA amplified using the polymerase chain reaction

Abstract

A 141-base pair fragment of human immunodeficiency virus-1 (HIV-1) DNA was amplified using the polymerase chain reaction (PCR). The products were slot-blotted onto a nitrocellulose membrane, revealed with a digoxigenin-labelled probe and quantitated by scanning densitometry. This method for the relative quantitation of HIV-1 DNA achieved reliable results and avoided the use of radioisotopes and the electrophoretic transfer of DNA. Testing of serial dilutions of HIV-1 extracted from infected cells revealed smooth titration curves. A reproducible increase in peripheral blood HIV-1 DNA was documented in a haemophilia patient during disease progression.