2017;31:1885C1889. mutation. Mutations conferring resistance to NRTIs and NNRTIs were present in 40.5% (123) and 69.1% (210), respectively. The most frequent mutations were K103N/S (48.0%), M184V (37.5%), and G190A/S (15.1%), and Y181C/G/V (14.1%). Expected drug resistance analysis exposed that 68.8% of the children experienced high-level resistance to NNRTIs and 11.5% had intermediate to high-level resistance to abacavir. Conclusions: This study showed high rates of resistance to NRTIs and NNRTIs among newly HIV-diagnosed children in Haiti, suggesting that in the era of option B+ (initiation of lifelong combination antiretroviral therapy to pregnant women with HIV), the majority of children who acquire HIV illness through MTCT have resistant HIV. These results possess led the National HIV System to revise the pediatric recommendations to include protease inhibitors in first-line regimens for those HIV-positive newborns. gene encompassing the protease and 5 section of the reverse transcriptase (RT) region was generated by RT-PCR and nested PCR. The purified PCR products were then sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, CA), and analyzed within the ABI Prism 3730 Genetic Analyzer (Applied Biosystems, Foster City, CA). The customized ReCALL software program was used to edit the uncooked sequences and generate consensus sequences15 and sequence quality assurance was performed on each newly obtained sequence using MEGA.16 HIVDR mutations and drug susceptibility profiles were identified using the HIVdb algorithm (version 8.4) deployed in the Stanford University or college Drug Resistance Database (http://hivdb.stanford.edu). Drug susceptibility profiles were interpreted such that the PF-3635659 presence of any drug resistance mutation that causes low-level, intermediate, or high-level of drug resistance was defined as resistance; those with vulnerable or potential low-level of resistance were designated as vulnerable. HIV-1 subtypes were identified using the REGA HIV subtyping tool.17 Statistical analyses The data were analyzed using SAS version 9.3 (SAS Institute, Cary, NC) and Epi Information 3.5.4 (CDC, Atlanta, 2013). Frequencies and chi-square checks were used to conclude categorical demographic data and mutation prevalence data while median and interquartile range [IQR] was reported for age. All graphics were produced using Microsoft Excel (Microsoft Corp., Redmond, WA, 2007). Honest considerations The study protocol was examined and authorized by the Haiti National Bioethics Committee and the Office of the Associate Director of Technology in the Center for Global Health in the Centers for Disease Control and Prevention. The study was identified to be not human being subjects study. Upon receiving the HIVDR results, the National HIV Program shared them with clinicians for patient management. RESULTS Geographic distribution and demographic characteristics of participants in the study Between January 1, 2013 and December 31, 2014, DBS samples collected from 3,555 HIV-exposed children from all 10 of Haitis geographic departments were submitted PF-3635659 to the LNSP for EID by PCR (Number 1). Of these, 360 (10.1%) were PCR-positive. Among the 360 HIV-positive DBS specimens, 355 experienced adequate residual DBS sample for inclusion in the study. Of the specimens submitted for genotyping, 304 (85.6%) were successfully genotyped, including 139 DBS samples collected in 2013 and 165 collected in 2014 (Number 1). The mean age of the children tested in 2013 was 6.8 months (standard deviation, SD 5.3 months), whereas the mean age of the children tested in 2014 was 6.2 months (S.D. 5.1 months); 243 (79.9%) of the children were under 6 months of age. Open in a separate window Number 1. Description of the study human population Prevalence of HIV-1 drug resistance mutations Among the 304 children for whom genotyping results were acquired, 217 (71.4%) had at least one DR mutation (Table 1), with 123 (40.5%) children having at least one DR mutation conferring resistance to nucleoside reverse transcriptase inhibitors (NRTIs) and 210 (69.1%) Rabbit polyclonal to ABHD12B having at least one DR mutation conferring resistance to non-NRTIs (NNRTIs). Moreover, 121 (39.8%) children harbored viruses with DR mutations conferring resistance to both NRTIs and NNRTIs, and 122 (40.1%) had two or more NNRTI mutations. Twenty-nine (9.5%) of the children had additional NNRTI mutations (A98G, E138A/G/K/Q,.Twenty-seven children had resistance to tenofovir: 5 had high-level resistance (18.5%), 9 had intermediate-level resistance (33.3%), and 13 had low-level resistance (48.1%). study showed high rates of resistance to NRTIs and NNRTIs among newly HIV-diagnosed children in Haiti, suggesting that in the era of PF-3635659 option B+ (initiation of lifelong combination antiretroviral therapy to pregnant women with HIV), the majority of children who acquire HIV contamination through MTCT have resistant HIV. These results have led the National HIV Program to revise the pediatric guidelines to include protease inhibitors in first-line regimens for all those HIV-positive newborns. gene encompassing the protease and 5 segment of the reverse transcriptase (RT) region was generated by RT-PCR and nested PCR. The purified PCR products were then sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, CA), and analyzed around the ABI Prism 3730 Genetic Analyzer (Applied Biosystems, Foster City, CA). The customized ReCALL software program was used to edit the natural sequences and generate consensus sequences15 and sequence quality assurance was performed on each newly obtained sequence using MEGA.16 HIVDR mutations and drug susceptibility profiles were decided using the HIVdb algorithm (version 8.4) deployed at the Stanford University or college Drug Resistance Database (http://hivdb.stanford.edu). Drug susceptibility profiles were interpreted such that the presence of any PF-3635659 drug resistance mutation that causes low-level, intermediate, or high-level of drug resistance was defined as resistance; those with susceptible or potential low-level of resistance were designated as susceptible. HIV-1 subtypes were decided using the REGA HIV subtyping tool.17 Statistical analyses The data were analyzed using SAS version 9.3 (SAS Institute, Cary, NC) and Epi Info 3.5.4 (CDC, Atlanta, 2013). Frequencies and chi-square assessments were used to summarize categorical demographic data and mutation prevalence data while median and interquartile range [IQR] was reported for age. All graphics were produced using Microsoft Excel (Microsoft Corp., Redmond, WA, 2007). Ethical considerations The study protocol was examined and approved by the Haiti National Bioethics Committee and the Office of the Associate Director of Science in the Center for Global Health at the Centers for Disease Control and Prevention. The study was determined to be not human subjects research. Upon receiving the HIVDR results, the National HIV Program shared them with clinicians for patient management. RESULTS Geographic distribution and demographic characteristics of participants in the study Between January 1, 2013 and December 31, 2014, DBS samples collected from 3,555 HIV-exposed children from all 10 of Haitis geographic departments were submitted to the LNSP for EID by PCR (Physique 1). Of these, 360 (10.1%) were PCR-positive. Among the 360 HIV-positive DBS specimens, 355 experienced sufficient residual DBS sample for inclusion in the study. Of the specimens submitted for genotyping, 304 (85.6%) were successfully genotyped, including 139 DBS samples collected in 2013 and 165 collected in 2014 (Physique 1). The mean age of the children tested in 2013 was 6.8 months (standard deviation, SD 5.3 months), whereas the mean age of the children tested in 2014 was 6.2 months (S.D. 5.1 months); 243 (79.9%) of the children were under 6 months of age. Open in a separate window Physique 1. Description of the study populace Prevalence of HIV-1 drug resistance mutations Among the 304 children for whom genotyping results were obtained, 217 (71.4%) had at least one DR mutation (Table 1), with 123 (40.5%) children having at least one DR mutation conferring resistance to nucleoside reverse transcriptase inhibitors.