Evaluation of the new Baxter MicroScan Automated Probe Analyzer for detection of HIV in clinical samples.

Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.

Evaluation of the new Baxter MicroScan Automated Probe Analyzer for detection of HIV in clinical samples.

Abstr Gen Meet Am Soc Microbiol. 1993;93:134 (abstract no. D220). Unique Identifier : AIDSLINE ASM93/93291787
Bush C; Sherman D; Donovan R; Peterson R; Godsey J; Henry Ford Hospital, Detroit, MI.

Abstract: A major limitation in the use of PCR in the clinical laboratory is the detection and quantitation of the amplified product. The limitations to current technology include the use of radioactive labels, labor intensive procedures, long time to results and lack of automation. This study applied the MicroScan Automated Probe Analyzer (MAPA) to the detection of HIV amplified by PCR in clinical samples. The PCR product was captured onto oligonucleotide coated beads and detected using a chlate-labeled secondary probe in a 10 minute sandwich-based hybridization assay carried out in a thermocycler. Beads from each assay were pipetted into a 96 well microtiter plate and placed into the MAPA instrument. The MAPA instrument processed the samples in 15 minutes by: 1) washing and vacuuming beads to remove excess probe, 2) dispensing terbium and incubating, and 3) determining the amount of signal in each assay well using a time-resolved fluorescence read. The MAPA detected HIV DNA from 78 of 78 PCR amplified peripheral blood mononuclear cell samples from HIV infected patients. The sensitivity of this technology was less than 100 copies when evaluated using purified HIV DNA. A plot of the log of DNA copies versus the log of time-resolved counts was linear between 10(2)-10(6) copies. A 32P evaluation of the same copy numbers of HIV gave similar results. In conclusion, the MAPA system overcomes the limitations of radioactive labels and lack of automation, and allows for determination of HIV positive samples in 30 minutes after amplification.

Keywords: Acquired Immunodeficiency Syndrome/*MICROBIOLOGY Autoanalysis/METHODS Comparative Study DNA, Viral/*ANALYSIS/GENETICS Human HIV/GENETICS/*ISOLATION & PURIF Nucleic Acid Hybridization Phosphorus Radioisotopes Polymerase Chain Reaction/METHODS Radioisotope Dilution Technique Virology/METHODS ABSTRACT
930930
M9391184

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