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Journal of Clinical Microbiology, April 2003, p. 1594-1599, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1594-1599.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Performance Characteristics of the TRUGENE HIV-1 Genotyping Kit and the Opengene DNA Sequencing System

Daniel R. Kuritzkes,1* Robert M. Grant,2 Paul Feorino,3 Marshal Griswold,4 Marie Hoover,5 Russell Young,1 Stephen Day,3 Robert M. Lloyd, Jr.,3 Caroline Reid,6 Gillian F. Morgan,6 and Dean L. Winslow6

Division of Infectious Diseases, University of Colorado Health Sciences Center, Denver, Colorado,1 Gladstone Institute of Virology and Immunology, San Francisco,2 Consolidated Laboratories, Van Nuys, California,4 Visible Genetics, Inc., Suwanee, Georgia,3 Advanced BioMedical Laboratories, Cinnaminson, New Jersey,5 Visible Genetics, Inc., Toronto, Ontario, Canada6

Received 11 February 2002/ Returned for modification 14 December 2002/ Accepted 9 January 2003

The TRUGENE HIV-1 Genotyping Kit and OpenGene DNA Sequencing System are designed to sequence the protease (PR)- and reverse transcriptase (RT)-coding regions of human immunodeficiency virus type 1 (HIV-1) pol. Studies were undertaken to determine the accuracy of this assay system in detecting resistance-associated mutations and to determine the effects of RNA extraction methods, anticoagulants, specimen handling, and potentially interfering substances. Samples were plasma obtained from HIV-infected subjects or seronegative plasma to which viruses derived from wild-type and mutant infectious molecular clones (IMC) of HIV-1 were added. Extraction methods tested included standard and UltraSensitive AMPLICOR HIV-1 MONITOR, QIAGEN viral RNA extraction mini kit, and QIAGEN Ultra HIV extraction kit, and NASBA manual HIV-1 quantitative NucliSens. Sequence data from test sites were compared to a "gold standard" reference sequence to determine the percent agreement. Comparisons between test and reference sequences at the nucleotide level showed 97.5 to 100% agreement. Similar results were obtained regardless of extraction method, regardless of use of EDTA or acid citrate dextrose as anticoagulant, and despite the presence of triglycerides, bilirubin, hemoglobin, antiretroviral drugs, HIV-2, hepatitis C virus (HCV), HBV, cytomegalovirus, human T-cell leukemia virus type 1 (HTLV-1), or HTLV-2. Samples with HIV-1 RNA titers of >=1,000 copies/ml gave consistent results. The TRUGENE HIV-1 Genotyping Kit and OpenGene DNA Sequencing System consistently generate highly accurate sequence data when tested with IMC-derived HIV and patient samples.

* Corresponding author. Present address: Partners AIDS Research Center, Brigham and Women’s Hospital, 65 Landsdowne St., Rm. 449, Cambridge, MA 02139. Phone: (617) 768-8371. Fax: (617) 768-8378. E-mail: dkuritzkes{at}partners.org.

Journal of Clinical Microbiology, April 2003, p. 1594-1599, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1594-1599.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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