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Transfusion. 2015 May;55(5):980-90. doi: 10.1111/trf.12936. Epub 2014 Nov 21.

Enhanced detection of viral diversity using partial and near full-length genomes of human immunodeficiency virus Type 1 provirus deep sequencing data from recently infected donors at four blood centers in Brazil.

Author information

1
Department of Virology, São Paulo Institute of Tropical Medicine, University of São Paulo, São Paulo, Brazil.
2
Clinical Laboratory, Department of Pathology, LIM 03, Hospital das Clínicas (HC), School of Medicine, University of São Paulo, São Paulo, Brazil.
3
Pernambuco State Center of Hematology and Hemotherapy-HEMOPE, Recife, Pernambuco, Brazil.
4
Hemorio, Rio de Janeiro, Brazil.
5
Minas Gerais State Center of Hematology and Hemotherapy-HEMOMINAS, Belo Horizonte, Minas Gerais, Brazil.
6
Department of Infectious Disease/Institute of Tropical Medicine, University of São Paulo, São Paulo, Brazil.
7
Blood Systems Research Institute, San Francisco, California.

Abstract

BACKGROUND:

Here, we report application of high-throughput near full-length genome (NFLG) and partial human immunodeficiency virus Type 1 (HIV-1) proviral genome deep sequencing to characterize HIV in recently infected blood donors at four major blood centers in Brazil.

STUDY DESIGN AND METHODS:

From 2007 to 2011, a total of 341 HIV+ blood donors from four blood centers were recruited to participate in a case-control study to identify HIV risk factors and motivations to donate. Forty-seven (17 from São Paulo, eight from Minas Gerais, 11 from Pernambuco, and 11 from Rio de Janeiro) were classified as recently infected based on testing by less-sensitive enzyme immunoassays. Five overlapping amplicons spanning the HIV genome were polymerase chain reaction amplified from peripheral blood mononuclear cells. The amplicons were molecularly barcoded, pooled, and sequenced by a paired-end protocol (Illumina).

RESULTS:

Of the 47 recently infected donor samples studied, 39 (82.9%) NFLGs and six (12.7%) partial fragments were de novo assembled into contiguous sequences and successfully subtyped. Subtype B was the only nonrecombinant virus identified in this study and accounted for 62.2% (28/45) of samples. The remaining 37.8% (17/45) of samples showed various patterns of subtype discordance in different regions of HIV-1 genomes, indicating two to four circulating recombinant subtypes derived from Clades B, F, and C. Fourteen samples (31.1%) from this study harbored drug resistance mutations, indicating higher rate of drug resistance among Brazilian blood donors.

CONCLUSION:

Our findings revealed a high proportion of HIV-1 recombinants among recently infected blood donors in Brazil, which has implications for future blood screening, diagnosis, therapy, and vaccine development.

PMID:
25413141
PMCID:
PMC4428985
DOI:
10.1111/trf.12936
[Indexed for MEDLINE]
Free PMC Article

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