6th International AIDS Conference San Francisco, California, USA — June 20-23, 1990

Int Conf AIDS 1990 Jun 20-23; 6:321 (abstract no. 1030)
Ruta M, Hewlett IK, Neiger N, Schumacher RT, Epstein J; DBBP, FDA, Bethesda, MD, USA

OBJECTIVE: To simultaneously detect HIV-1 and HBV DNA in clinical specimens by co-amplification using PCR.

METHODS: Plasmids carrying HIV-1 and HBV sequences served as templates for optimization of the co-amplification reaction. HBV primers to the core region and conditions for amplification were as described by S.Kaneko et al., (J.C.M. 1989, 27; 1930-1933). A single series of HBV amplifications using primers 1763 and 2032R was compared to the secondary amplification using nested primers 1778E and 2017RB. Following amplification, the products were detected by liquid hybridization using an internally derived 32-P-labelled oligonucleotide (1820). For HIV detection, primers to the gag region SK38/39 and an internally derived primer SK19 were used. Products were analyzed by PAGE followed by autoradiography. DNA extracted from the serum of 2 patients with co-infections and 3 uninfected individuals was analyzed to establish utility of the assay on clinical samples.

RESULTS: Co-amplification of these viral sequences resulted in a limit of detection of 10 copies of the HIV-1 and less than 500 copies of the HBV genome. By this method, 2/2 patients with co-infections tested positive for both viruses (DNA) by PCR. In the same assay, none of three infected individuals had evidence of viral sequences in their serum.

CONCLUSIONS: We have developed a simple and specific PCR-based assay for simultaneous detection of HIV-1 and HBV DNA in the serum of infected individuals. This assay may be of value in the analysis of clinical samples and in determining the role of co-factors in disease progression.

900620
1030

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