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.
Subject(s)
Blood/virology , Drug Resistance, Viral/genetics , HIV-1/drug effects , Microbial Sensitivity Tests/methods , Specimen Handling/methods , Amino Acid Substitution/genetics , Genotype , HIV-1/genetics , Humans , Mutation, Missense , Plasma/virology , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Prevalence studies indicate that transmission of drug-resistant HIV has been rising in the adult population, but data from the perinatally infected pediatric population are limited. In this retrospective study, we sequenced the pol region of HIV from perinatally infected infants diagnosed in New York State in 2001-2002. Analyses of drug resistance, subtype diversity, and perinatal antiretroviral exposure were conducted, and the results were compared with those from a previous study of HIV-infected infants identified in 1998-1999. Eight of 42 infants (19.1%) had provirus carrying at least 1 drug-resistance mutation, an increase of 58% over the 1998-1999 results. Mutations conferring resistance to nucleoside reverse transcriptase inhibitors, nonnucleoside reverse transcriptase inhibitors, and protease inhibitors were detected in 7.1%, 11.9%, and 2.4% of specimens, respectively. Consistent with previous results, perinatal antiretroviral exposure was not associated with drug resistance (P = 0.70). Phylogenetic analysis indicated that 16.7% of infants were infected with a non-subtype B strain of HIV. It seems that drug-resistant and non-subtype B strains of HIV are becoming increasingly common in the perinatally infected population. Our results highlight the value of resistance testing for all HIV-infected infants upon diagnosis and the need to consider subtype diversity in diagnostic and treatment strategies.
Subject(s)
Anti-HIV Agents/pharmacology , HIV Infections/virology , HIV/drug effects , HIV/genetics , Mutation , Anti-HIV Agents/therapeutic use , Cluster Analysis , DNA Mutational Analysis , Drug Resistance, Viral/genetics , Female , Genome, Viral , Genotype , HIV/classification , HIV Infections/drug therapy , HIV Infections/transmission , Humans , Infant, Newborn , Infectious Disease Transmission, Vertical , New York , Phylogeny , Pregnancy , Pregnancy Complications, Infectious/drug therapy , Pregnancy Complications, Infectious/virology , RNA, Viral/genetics , Retrospective Studies , Sequence Analysis, DNA , Sequence Homology , Statistics as TopicABSTRACT
OBJECTIVE: To assess trends in HIV, hepatitis C virus (HCV) and HIV/HCV infection among injecting drug users (IDU) from 1990 to 2001 in New York City. The 1990-2001 time period included a very large expansion of syringe exchange in New York City, from 250,000 to 3,000,000 syringes exchanged annually. METHODS: Cross-sectional seroprevalence surveys of IDU entering drug abuse treatment in New York City, with sample sizes for HCV of 72 in 1990-1991 and 412 in 2000-2001. A structured risk behavior questionnaire was administered, and HIV and HCV testing were conducted. HCV testing was performed on de-linked stored serum samples. RESULTS: Over the 1990-2001 period, HIV prevalence declined from 54 to 13%. HCV prevalence declined from 80 to 59% among HIV-seronegative individuals, and from 90 to 63% overall. The estimated HCV incidence in 2000-2001 among new injectors was 18 per 100 person-years at risk. CONCLUSIONS: The large-scale expansion of syringe exchange was temporally associated with large reductions in both HIV and HCV prevalence. The prevalence and incidence of HCV, however, still remain at high levels among IDU in New York City.
