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Clinical and Vaccine Immunology, February 2006, p. 235-238, Vol. 13, No. 2
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.2.235-238.2006

Performance of the TruGene Human Immunodeficiency Virus Type 1 Genotyping Kit and OpenGene DNA Sequencing System on Clinical Samples Diluted to Approximately 100 Copies per Milliliter

Howard B. Gale,* Virginia L. Kan, and Rebecca C. Shinol

Infectious Diseases Laboratory, Infectious Diseases Section, Medical Service, Veterans Affairs Medical Center, Washington, D.C

Received 25 July 2005/ Returned for modification 14 November 2005/ Accepted 8 December 2005

The TruGene human immunodeficiency virus type 1 (HIV-1) genotyping kit/OpenGene DNA sequencing system (Bayer HealthCare, Tarrytown, NY) reliably produced clinically acceptable resistance profiles for reverse transcriptase and protease inhibitors on patient samples diluted to ~100 copies/ml following extraction with the QIAamp viral RNA minikit (QIAGEN Inc., Valencia, CA). One modification of the standard protocol was made to guarantee PCR amplification: a centrifugation step to concentrate virus was added before RNA extraction. For genotypic antiretroviral resistance testing, no significant differences in the identification and sensitivity of detection for codon mutations, base mutations, and multibase sites were found between the original and diluted samples.

* Corresponding author. Mailing address: Infectious Diseases Section (151B), Veterans Affairs Medical Center, 50 Irving Street N.W., Washington, DC 20422. Phone: (202) 745-8000, ext. 7998. Fax: (202) 745-8432. E-mail: howard.gale{at}med.va.gov.

Clinical and Vaccine Immunology, February 2006, p. 235-238, Vol. 13, No. 2
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.2.235-238.2006




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