HIV-1 RNA testing of pooled dried blood spots is feasible to diagnose acute HIV infection in resource limited settings

Objectives:Rapid human immunodeficiency virus (HIV) antibody tests, routinely used for diagnosis in adults and older children in resource-limited settings (RLS), do not detect early HIV infections prior to seroconversion or when antibody levels are still low. Nucleic acid amplification to detect HIV-1 RNA is the most sensitive method for acute HIV infection diagnosis, but is costly. We therefore investigated HIV- 1 RNA testing of pooled dried blood spots (DBS) to diagnose acute HIV infection.Design:Laboratory-based investigation.Methods:DBS were collected from HIV-1 Voluntary Counselling and Testing (HVCT) clients who tested negative on the Advanced QualityTM HIV antibody rapid test. DBS samples from five participants were pooled and tested on the COBAS AmpliPrep/COBAS TaqMan HIV-1 (CAP/CTM) Test v2. Individual DBS were tested when pools tested positive ( 200 RNA copies/ml). Acute infection was confirmed by HIV viral load testing, two fourth-generation HIV serological assays, and Geenius™ HIV 1/2 Assay for antibody band identification.Results:Of 482 participants who were tested, one (0.2%) had acute. HIV infection: Fourth generation serology was low-level positive, the plasma HIV viral load was 15 929 HIV-1 RNA copies/ml, gp160 and gp41 antibody bands were positive and the p31 band was negative, indicating a Fiebig Stage 5 infection.Conclusions: Pooled DBS HIV-1 RNA testing is efficient compared to individual testing for acute HIV infection diagnosis. Early identification of participants with acute HIV infection facilitates immediate initiation of antiretroviral therapy to improve immune recovery and prevent transmission to others.

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Use of Dried Blood Spots to Elucidate Full-Length Transmitted/Founder HIV-1 Genomes

Pathogens and Immunity ◽

10.20411/pai.v1i1.116 ◽

2016 ◽

Vol 1 (1) ◽

pp. 129 ◽

Cited By ~ 4

Author(s):

Jesus F. Salazar-Gonzalez ◽

Maria G. Salazar ◽

Damien C. Tully ◽

Colin B. Ogilvie ◽

Gerald H. Learn ◽

...

Keyword(s):

Dried Blood Spots ◽

Full Length ◽

Cold Chain ◽

Clinical Samples ◽

Envelope Gene ◽

Acid Extraction ◽

Resource Limited Settings ◽

Blood Spots ◽

Resource Limited ◽

Hiv 1

Background: Identification of HIV-1 genomes responsible for establishing clinical infection in newly infected individuals is fundamental to prevention and pathogenesis research. Processing, storage, and transportation of the clinical samples required to perform these virologic assays in resource-limited settings requires challenging venipuncture and cold chain logistics. Here, we validate the use of dried-blood spots (DBS) as a simple and convenient alternative to collecting and storing frozen plasma.Methods: We performed parallel nucleic acid extraction, single genome amplification (SGA), next generation sequencing (NGS), and phylogenetic analyses on plasma and DBS.Results: We demonstrated the capacity to extract viral RNA from DBS and perform SGA to infer the complete nucleotide sequence of the transmitted/founder (TF) HIV-1 envelope gene and full-length genome in two acutely infected individuals. Using both SGA and NGS methodologies, we showed that sequences generated from DBS and plasma display comparable phylogenetic patterns in both acute and chronic infection. SGA was successful on samples with a range of plasma viremia, including samples as low as 1,700 copies/ml and an estimated ~50 viral copies per blood spot. Further, we demonstrated reproducible efficiency in gp160 env sequencing in DBS stored at ambient temperature for up to three weeks or at -20ºC for up to five months.Conclusions: These findings support the use of DBS as a practical and cost-effective alternative to frozen plasma for clinical trials and translational research conducted in resource-limited settings.

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Early HIV-1 Diagnosis Using In-House Real-Time PCR Amplification on Dried Blood Spots for Infants in Remote and Resource-Limited Settings

JAIDS Journal of Acquired Immune Deficiency Syndromes ◽

10.1097/qai.0b013e31818e2531 ◽

2008 ◽

Vol 49 (5) ◽

pp. 465-471 ◽

Cited By ~ 26

Author(s):

Nicole Ngo-Giang-Huong ◽

Woottichai Khamduang ◽

Baptiste Leurent ◽

Intira Collins ◽

Issaren Nantasen ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Pcr Amplification ◽

Dried Blood Spots ◽

Resource Limited Settings ◽

Blood Spots ◽

Resource Limited ◽

Hiv 1

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IP-10 Levels as an Accurate Screening Tool to Detect Acute HIV Infection in Resource-Limited Settings

Scientific Reports ◽

10.1038/s41598-017-08218-0 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 9

Author(s):

Lucía Pastor ◽

Aina Casellas ◽

Jorge Carrillo ◽

Sergi Alonso ◽

Erica Parker ◽

...

Keyword(s):

Hiv Infection ◽

Screening Tool ◽

Resource Limited Settings ◽

Acute Hiv Infection ◽

Resource Limited ◽

Acute Hiv

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Evaluation of a manual DNA extraction protocol and an isothermal amplification assay for detecting HIV-1 DNA from dried blood spots for use in resource-limited settings

Journal of Clinical Virology ◽

10.1016/j.jcv.2012.01.004 ◽

2012 ◽

Vol 54 (1) ◽

pp. 11-14 ◽

Cited By ~ 12

Author(s):

Jeanne A. Jordan ◽

Christine O. Ibe ◽

Miranda S. Moore ◽

Christel Host ◽

Gary L. Simon

Keyword(s):

Dna Extraction ◽

Dried Blood Spots ◽

Isothermal Amplification ◽

Resource Limited Settings ◽

Blood Spots ◽

Extraction Protocol ◽

Resource Limited ◽

Amplification Assay ◽

Hiv 1

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Decreased Seroreactivity in Individuals Initiating Antiretroviral Therapy during Acute HIV Infection

Journal of Clinical Microbiology ◽

10.1128/jcm.00757-19 ◽

2019 ◽

Vol 57 (10) ◽

Cited By ~ 10

Author(s):

Mark M. Manak ◽

Linda L. Jagodzinski ◽

Ashley Shutt ◽

Jennifer A. Malia ◽

Mike Leos ◽

...

Keyword(s):

Antiretroviral Therapy ◽

Hiv Infection ◽

Western Blot ◽

Research Collaboration ◽

Virologic Suppression ◽

Infection Status ◽

Acute Hiv Infection ◽

Western Blot Assay ◽

Acute Hiv ◽

Hiv 1

ABSTRACTAntiretroviral therapy (ART) during acute HIV infection (AHI) interrupts viral dynamics and may delay the emergence of serological markers targeted by current HIV screening and confirmatory assays, thus creating challenges for correctly classifying HIV infection status. The performance of three HIV antigen/antibody combination (HIV Ag/Ab Combo) assays (the Bio-Rad GS, Abbott Architect, and Bio-Rad BioPlex 2200 assays) was evaluated with samples collected from RV254/South East Asia Research Collaboration in HIV 010 (RV254/SEARCH010) study (Bangkok, Thailand) participants at weeks 12 and 24 following the initiation of ART at Fiebig stage I (FI) (n = 23), FII (n = 39), or FIII/IV (n = 22). Supplemental, confirmatory testing was performed by the Geenius HIV 1/2 and HIV-1 Western blot assays (Bio-Rad). Samples from 30 untreated, HIV-1-infected individuals demonstrated robust HIV Ag/Ab Combo assay reactivity with well-developed HIV-1 Western blotting profiles by 24 weeks after infection. In contrast, 52.2% of samples from individuals initiating ART at FI, 7.7% of samples from individuals initiating ART at FII, and 4.5% of samples from individuals initiating ART at FIII/IV were nonreactive by the HIV Ag/Ab Combo assays, with 36.4 to 39.1% of samples having low signal-to-cutoff (S/CO) results by the Architect and BioPlex assays (S/CO < 10). Seroreversion from a reactive to a nonreactive status was observed in 10 individuals initiating ART at FII and 3 individuals initiating ART at FIII/IV. The Geenius and HIV-1 Western blot assay results were negative or indeterminate for 73.9% and 69.6% of individuals, respectively, treated at FI; 50.0% and 26.3% of individuals, respectively, treated at FII; and 54.5% and 40.9% of individuals, respectively, treated at FIII/IV. Virologic suppression of HIV-1 by ART during AHI impedes seroconversion to biomarkers of infection, limiting the utility of HIV Ag/Ab Combo and supplemental, confirmatory assays for infection status determination.

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