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Journal of Clinical Microbiology, June 1998, p. 1534-1538, Vol. 36, No. 6
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Detection of Human Immunodeficiency Virus Type 1 (HIV-1) RNA in Pools of Sera Negative for Antibodies to HIV-1 and HIV-2

Pierre-Alain Morandi,1 Gérard A. Schockmel,1 Sabine Yerly,1 Philippe Burgisser,2 Peter Erb,3 Lukas Matter,4 Radan Sitavanc,5 and Luc Perrin1,*

Laboratory of Virology and AIDS Center, Division of Infectious Diseases, Geneva University Hospital, 1211 Geneva,1 Division of Immunology and Allergy, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne,2 Institute for Medical Microbiology, University of Basel, 4003 Basel,3 Institute for Medical Microbiology, University of Bern, 3010 Bern,4 and Bio Analytique Institute, 1207 Geneva,5 Switzerland

Received 22 December 1997/Returned for modification 25 February 1998/Accepted 17 March 1998

A total of 234 pools were prepared from 10,692 consecutive serum samples negative for antibodies to human immunodeficiency virus type 1 (HIV-1) and HIV-2 collected at five virological laboratories (average pool size, 45 serum samples). Pools were screened for the presence of HIV-1 RNA by a modified commercial assay (Amplicor HIV-1 Monitor test) which included an additional polyethylene glycol (PEG) precipitation step prior to purification of viral RNA (PEG Amplicor assay). The sensitivity of this assay for HIV-1 RNA detection in individual serum samples within pools matches that of standard commercial assays for individual serum samples, i.e., 500 HIV-1 RNA copies per ml. Five pools were identified as positive, and each one contained one antibody-negative, HIV-1 RNA-positive serum sample, corresponding to an average of 1 infected sample per 2,138 serum samples. Retrospective analysis revealed that the five HIV-1 RNA-positive specimens originated from individuals who had symptomatic primary HIV-1 infection at the time of sample collection and who were also positive for p24 antigenemia. We next assessed the possibility of performing the prepurification step by high-speed centrifugation (50,000 × g for 80 min) of 1.5-ml pools containing 25 µl of 60 individual serum samples, of which only 1 contained HIV-1 RNA (centrifugation Amplicor assay). The sensitivity of this assay also matches the sensitivities of standard commercial assays for HIV-1 RNA detection in individual serum samples. The results demonstrate that both assays with pooled sera can be applied to the screening of large numbers of serum samples in a time- and cost-efficient manner.

* Corresponding author. Mailing address: Laboratory of Virology and AIDS Center, Division of Infectious Diseases, Geneva University Hospital, 1211 Geneva 14, Switzerland. Phone: 41.22/37.24.991. Fax: 41.22/37.24.990. E-mail: luc.perrin{at}hcuge.ch.

Journal of Clinical Microbiology, June 1998, p. 1534-1538, Vol. 36, No. 6
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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Copyright © 1998 by the American Society for Microbiology. All rights reserved.