Evaluation of dried blood spot specimens for HIV-1 drug-resistance testing using the Trugene® HIV-1 genotyping assay

• Published in: Journal of clinical virology 2008-04, Vol.41 (4), p.283-287
• Main Authors: Hallack, Renee, Doherty, Lauren E, Wethers, Judith A, Parker, Monica M
• Format: Article
• Language: English
• Subjects: Allergy and Immunology; Amino Acid Substitution - genetics; Biological and medical sciences; Blood - virology; Dried blood spot; Drug resistance; Drug Resistance, Viral - genetics; Fundamental and applied biological sciences. Psychology; Genotype; Genotyping; HIV-1; HIV-1 - drug effects; HIV-1 - genetics; Human viral diseases; Humans; Infectious Disease; Infectious diseases; Medical sciences; Microbial Sensitivity Tests - methods; Microbiology; Miscellaneous; Mutation, Missense; Plasma - virology; Reproducibility of Results; Sensitivity and Specificity; Specimen Handling - methods; Trugene ® HIV-1 genotyping assay; Viral diseases; Viral diseases of the lymphoid tissue and the blood. Aids; Virology
• ISSN: 1386-6532; 1873-5967
• DOI: 10.1016/j.jcv.2007.12.011

Abstract Background Efforts to simplify the collection and shipping of specimens for HIV drug-resistance testing in resource-limited settings are needed as antiretroviral therapy increases worldwide. Objective To evaluate the reliability and practicality of using dried blood spots (DBS) for HIV-1 drug-resistance testing with the Trugene® HIV-1 genotyping assay. Study design Nucleic acids from 33 DBS and counterpart plasma specimens were extracted using the Nuclisens® MiniMAG™ system and genotyped using the Trugene® HIV-1 genotyping assay. Results were evaluated for sensitivity, accuracy, and reproducibility. Results A genotype was obtained for 33 (100%) plasma specimens and 26 (78.8%) DBS specimens, including 19 of 21 (90.5%) DBS specimens with a viral load greater than 6000 copies/mL. The mean nucleotide sequence concordance for the 940-nucleotide region evaluated was 99.3% for 26 DBS and plasma pairs, and 99.2% for 15 replicate DBS pairs. All 58 resistance-associated mutations detected in plasma specimens were detected in the corresponding DBS specimens. Conclusions We show that DBS can be reliably and accurately genotyped using standard clinical assay methods, offering a practical alternative to plasma. This method is well suited for pre-treatment resistance testing and has potential for use in monitoring drug resistance in ART-treated individuals.