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Journal of Clinical Microbiology, February 2000, p. 716-723, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Performance of a Multiplex Qualitative PCR LCx Assay for Detection of Human Immunodeficiency Virus Type 1 (HIV-1) Group M Subtypes, Group O, and HIV-2

Klara Abravaya,* Claudia Esping, Robert Hoenle, Jacek Gorzowski, Robert Perry, Paul Kroeger, John Robinson, and Richard Flanders

Abbott Laboratories, Abbott Park, Illinois 60064

Received 20 May 1999/Returned for modification 16 September 1999/Accepted 11 November 1999

Early detection of human immunodeficiency virus (HIV) in blood and blood products can be achieved by a sensitive nucleic acid amplification-based assay. We report on the performance of a PCR-based qualitative assay that detects both HIV type 1 (HIV-1) and HIV-2 with a sensitivity of 20 to 50 copies/ml. The assay has a specificity of 99.6% and an inhibition rate of 1.7%. One milliliter of sample is processed with a manifold system and Qiagen columns, and one-third of the extracted sample is used for PCR amplification. An internal control sequence, which is processed and amplified with each sample, monitors for amplification inhibition. Samples are reverse transcribed and are then amplified by reverse transcription-coupled PCR, after which HIV-1- and HIV-2-specific probes are hybridized to the amplified products. Following hybridization, samples are detected in the LCx instrument by microparticle enzyme immunoassay techniques. The detection system has an automated inactivation step that controls for PCR contamination. The HIV-1/2 qualitative RNA assay detects HIV-1 group M subtypes A, B, C, D, E, F, and G and group O. Testing of several HIV-1 seroconversion panels has demonstrated that the HIV-1/2 qualitative RNA assay detects HIV infection on the average of 6 days before p24 antigen can be detected and 11 days before antibodies can be detected.

* Corresponding author. Mailing address: Abbott Laboratories, Dept. 09ND/AP20, 100 Abbott Park, Abbott Park, IL 60064-3500. Phone: (847) 938-1410. Fax: (847) 938-8777. E-mail: klara.abravaya{at}add.ssw.abbott.com.

Journal of Clinical Microbiology, February 2000, p. 716-723, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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