Quest Diagnostics Investments, Inc.

Jamie L Platt

Maurice Exner

Erik P Johnson

Dale A Schwab

Nik Isely

Martin Siaw

Ron M Kagen

Described herein are methods for direct detection of microbial agent(s) in a polymicrobial sample, such as a biological sample from a human, without culturing the microbial agent(s). The direct detection can identify mixtures of bacteria and/or fungi in the sample. Also described are primer sequences and amplification techniques for performing the direct detection methods.

Method for direct microbial identification

Disclosed is a method for detecting human retroviral nucleic acids such as human immunodeficiency virus type I (HIV-1) nucleic acid, human T-cell leukemia virus type I (HTLV-I) nucleic acid, and human T-cell leukemia virus type II (HTLV-II) nucleic acid in a sample. In the method, the sample is treated with reverse transcriptase to generate cDNA, and the cDNA is subsequently analyzed to detect HIV-1, HTLV-I, and HTLV-II. The method may include performing PCR and the method may utilize specific primers. In addition, the method may utilize HTLV linkers that facilitate PCR amplification and sequencing. The cDNA may be treated with restriction enzymes before or after PCR amplification to facilitate sequencing and detection of HIV-1, HTLV-I, or HTLV-II nucleic acid.

Methods for detecting retroviruses

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San Juan Capistrano, CA, United States of America

Jamie L Platt