The qualitative Roche HIV-1 DNA Amplicor assay continues to be used
The qualitative Roche HIV-1 DNA Amplicor assay continues to be used for days gone by twenty years to diagnose HIV infection in infants and small children but has been phased out; therefore, alternative assays should be discovered. CI, 97.2 to 99.9%). These total results claim that the assay would work for early infant diagnosis of HIV-1. Launch The Joint US Plan on HIV/Helps (UNAIDS) quotes that children significantly less than 15 years of age accounted for roughly 13% of fresh HIV infections in 2011 (1). Additionally, a meta-analysis concluded that with no treatment, about 35% of African HIV-infected children pass away before their 1st birthday, and more than 50% pass away by the time they may be 2 years older (2). However, data from the Children with HIV Early Antiretroviral Therapy (CHER) trial shown that early, as opposed to delayed, initiation of antiretrovirals (ARVs) in young infants significantly reduced mortality (3), and data from several clinical trials possess indicated that infected infants exposed to prophylactic ARVs often develop resistance to the medicines, which may limit future restorative drug choices (4C6). Therefore, early recognition of HIV illness in infants Bupivacaine HCl supplier is important so that ARV prophylaxis can be stopped to reduce the development of resistance to the ARVs, and therapeutic doses of ARVs can be initiated as quickly as Bupivacaine HCl supplier possible. Early diagnosis of HIV infection using serologic testing of antibodies, however, is hindered by the presence of maternal antibodies that cross the placenta during gestation. Consequently, early infant diagnostics must test for either viral antigens or nucleic acids. For many years, the Roche Amplicor HIV-1 DNA assay, version 1.5, has been the mainstay and gold standard for early infant diagnosis (EID), having been validated and used extensively in many countries and recommended by the World Health Organization, as well as the U.S. Centers for Disease Control and Prevention, for EID programs using both whole-blood pellets and dried blood spots (DBS) (7). However, Roche plans to discontinue this assay in the next few years, so alternative assays must be found (8). Although there are several alternatives, including the Roche TaqMan and Abbott RealTime quantitative HIV-1 RNA and Rabbit polyclonal to ACAD8 qualitative HIV-1 total nucleic acid assays, the Gen-Probe Aptima HIV-1 RNA qualitative assay is the only nucleic acid assay currently approved by the FDA for HIV diagnosis using serum or plasma (9). Although its utility in EID using DBS (10C12) and whole blood (11) has been reported, data on the use of this assay using infant plasma are limited. The New York State Department of Health evaluated Aptima for infant diagnosis but tested only plasma from 28 HIV-exposed uninfected babies and 68 HIV-infected infants (T. Bupivacaine HCl supplier J. Sullivan, T. T. Miller, B. Warren, M. M. Parker, presented at the Third HIV Diagnostics Conference, Orlando, FL, 24 to 26 March 2010). An additional 48 sera from HIV-exposed, uninfected infants were tested and found to be nonreactive in the Aptima assay (13). MATERIALS AND METHODS Samples. The limit of detection, within-run repeatability, and between-run reproducibility were assessed using control material obtained from the Virus Quality Assurance Program (VQA; Rush University Medical Center, Chicago, IL) (14) diluted in Basematrix (SeraCare, Milford, MA). Repeatability was assessed by diluting the VQA HIV-1 RNA 200-copy/ml (cp/ml) control material to final concentrations of 100 and 25 HIV-1 RNA cp/ml and testing in duplicate in 4 separate runs by two technologists. Reproducibility was determined by testing three positive VQA controls (10,000, 200, and 50 cp/ml) and a negative control in singlet over 5 days using 2 different kit lots. To assess the limit of detection, we made additional dilutions to 0.4 cp/ml that were tested in 4 separate runs by two technologists using 3 kit lots. Fewer samples were tested at the highest concentrations (= Bupivacaine HCl supplier 5 to 8 replicates) than at lower concentrations, which were tested more frequently (= 10 to 20 replicates). There were a few invalid results that were not included in the analysis. Basematrix was used as HIV RNA negative-control material for all.