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Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots

dc.contributor.authorMartin, Francisco
dc.contributor.authorPalladino, Claudia
dc.contributor.authorMateus, Rita
dc.contributor.authorBolzan, Anna
dc.contributor.authorGomes, Perpétua
dc.contributor.authorBrito, José
dc.contributor.authorCarvalho, Ana Patrícia
dc.contributor.authorCardoso, Yolanda
dc.contributor.authorDomingos, Cristovão
dc.contributor.authorClemente, Vanda Sofia Lôa
dc.contributor.authorTaveira, Nuno
dc.date.accessioned2019-11-21T12:37:30Z
dc.date.available2019-11-21T12:37:30Z
dc.date.issued2017-06
dc.descriptionThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.pt_PT
dc.description.abstractBACKGROUND: Early diagnosis and treatment reduces HIV-1-related mortality, morbidity and size of viral reservoirs in infants infected perinatally. Commercial molecular tests enable the early diagnosis of infection in infants but the high cost and low sensitivity with dried blood spots (DBS) limit their use in sub-Saharan Africa. OBJECTIVES: To develop and validate a sensitive and cheap qualitative proviral DNA PCR-based assay for early infant diagnosis (EID) in HIV-1-exposed infants using DBS samples. STUDY DESIGN: Chelex-based method was used to extract DNA from DBS samples followed by a nested PCR assay using primers for the HIV-1 integrase gene. Limit of detection (LoD) was determined by Probit regression using limiting dilutions of newly produced recombinant plasmids with the integrase gene of all HIV-1 subtypes and ACH-2 cells. Clinical sensitivity and specificity were evaluated on 100 HIV-1 infected adults; 5 infected infants; 50 healthy volunteers; 139 HIV-1-exposed infants of the Angolan Pediatric HIV Cohort (APEHC) with serology at 18 months of life. RESULTS: All subtypes and CRF02_AG were amplified with a LoD of 14 copies. HIV-1 infection in infants was detected at month 1 of life. Sensitivity rate in adults varied with viral load, while diagnostic specificity was 100%. The percentage of HIV-1 MTCT cases between January 2012 and October 2014 was 2.2%. The cost per test was 8-10 USD which is 2- to 4-fold lower in comparison to commercial assays. CONCLUSIONS: The new PCR assay enables early and accurate EID. The simplicity and low-cost of the assay make it suitable for generalized implementation in Angola and other resource-constrained countries.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMartin F, Palladino C, Mateus R, Bolzan A, Gomes P, Brito J, et al. (2017) Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots. PLoS ONE 12(7): e0181352. https://doi.org/10.1371/journal.pone.0181352pt_PT
dc.identifier.doi10.1371/journal.pone.0181352pt_PT
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/10400.26/30243
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherPLoSpt_PT
dc.relation.publisherversionhttps://doi.org/10.1371/journal.pone.0181352pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectHIV-1 infectionpt_PT
dc.subjectInfant diagnosispt_PT
dc.subjectDNA PCR assaypt_PT
dc.subjectAngolapt_PT
dc.subjectAngolapt_PT
dc.subjectDried blood spotspt_PT
dc.titleEarly infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spotspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.startPagee0181352pt_PT
oaire.citation.titlePLoS ONEpt_PT
oaire.citation.volume12(7)pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT

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