Detection and characterization of HIV-1 by polymerase chain reaction

Abstract

Recently a new technique, the polymerase chain reaction (PCR), has been used for the detection and characterization of HIV-1 proviral DNA and viral RNA. These reports support the notion that the PCR is more sensitive and specific than other established HIV-1 detection techniques. However, due to its extreme sensitivity, the PCR is highly susceptible to contamination, resulting in false positive results. To avoid contamination, strict rules on sample preparation and pre- and post-PCR handling are required. Confirmation of both positive and negative PCR results by independent techniques is not always feasible, and, therefore, optimal PCR conditions, inclusion of control samples, repetition of results, and confirmation of specificity by hybridization are required. The choice of the material from which HIV-1 is amplified, the primers used for amplification as well as the PCR conditions will determine what is actually amplified.