POLYMERASE CHAIN-REACTION DETECTION OF HUMAN-IMMUNODEFICIENCY-VIRUS DNA IN HUMAN PERIRADICULAR LESIONS

Human immunodeficiency virus (HIV) has been previously reported to be present in the dental pulp of a patient with AIDS. The present report investigated the feasibility of polymerase chain reaction (PCR) amplification to detect the HIV proviral DNA in cells from periradicular lesions from an HIV-positive patient. The standard PCR amplification with 30 cycles and the nested PCR consisting of two 25-cycle amplifications were used. Samples from each reaction were separated by nondenaturing polyacrylamide gel electrophoresis and stained with ethidium bromide for visualization. Gels were electroblotted to nylon membranes, which were then fixed, denatured, and dried. Membranes were hybridized to specific radioactive oligonucleotide probes and placed next to Kodak XAR film for visualization of the HIV-specific bands. No evidence of HIV-specific reaction was observed in cells (negative control) or in two periradicular lesions from two HIV-negative patients. The ethidium bromide strains revealed that PCR amplification of DNA extracts from two lesions from the HIV-positive patient yielded PCR bands (with both primer pairs) which corresponded to HIV-specific bands of the expected size.