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

Abstract

Background In resource-limited settings, most perinatally HIV-1-infected infants do not receive timely antiretroviral therapy because early HIV-1 diagnosis is not available or affordable. Objective To assess the performance of a low cost in-house real-time PCR assay to detect HIV-1 DNA in infant dried blood spots (DBS). Methods 1319 DBS collected throughout Thailand from non-breastfed infants born to HIV-1-infected mothers were shipped at room temperature to a central laboratory. In-house real-time DNA-PCR results were compared to Roche Amplicor® HIV-1 DNA test (Version 1.5) results. In addition, we verified the Roche test performance on DBS sampled from 1218 other infants using as reference HIV serology result at 18 months of age. Results Real-time DNA-PCR and Roche DNA-PCR results were 100% concordant. Compared to HIV-serology results, the Roche test sensitivity was 98.6% (95% CI: 92.6 to 100.0%) and its specificity at 4 months of age was 99.7% (95% CI: 99.2 to 99.9%). Conclusions In-house real-time PCR performed as well as the Roche test in detecting HIV-1 DNA on DBS in Thailand. Combined use of DBS and real-time PCR assays is a reliable and affordable tool to expand access to early HIV-1 diagnosis in remote and resource-limited settings, enabling timely treatment for HIV-1-infected infants.

Long-Term Survival of HIV-Infected Children Receiving Antiretroviral Therapy in Thailand: A 5-Year Observational Cohort Study

“…Furthermore, with the expansion of access to early HIV diagnosis [34][35][36], many countries have reported challenges in increasing uptake among HIV-exposed infants and retaining newly diagnosed infants [37,38]. This highlights the need for strengthening counselling services for new mothers and referral linkages between prevention of mother-to-child transmission of HIV and treatment programs.…”

Evaluation of blood collection filter papers for HIV-1 DNA PCR

“…Therefore, DBS offers several advantages in areas that lack appropriate infrastructure for processing blood and cold-chain for transportation and storage of blood products. [28][29][30]32,33,36 Most HIV-1 DNA testing has been performed on Schleicher & Schuell 903 (or Whatman 903, W-903 (Whatman Inc., Florham Park, NJ, USA)) paper, [5][6][7][8][9][10][11]16,18,19,28,30,31,34,36,38 but limited data are available for other types of blood collection paper, 2,17,27,29,32,35,37 including Ahlstrom grade 226 (ID Biological Systems, Greenville, SC, USA) and Munktell TFN (Munktell Filter AB, Falun, Sweden). These collection papers are manufactured to meet the CLSI (Clinical and Laboratory Standards Institute) standard for Blood Collection on Filter Paper for Newborn Screening programs (LA4-A5, vol.…”

“…[26][27][28][29][30] Qualitative HIV-1 DNA detection has been performed on DBS using commercial and in-house molecular assays, and it is the preferred method for early infant diagnosis of HIV (EID) in RLS. 18,19,[31][32][33][34][35][36][37][38] DBS can be collected heel or finger prick and without the need for venipuncture, stored at RT, and more easily transported to the testing laboratory at RT and with less biohazard risk. Therefore, DBS offers several advantages in areas that lack appropriate infrastructure for processing blood and cold-chain for transportation and storage of blood products.…”

RNA versus DNA (NucliSENS EasyQ HIV-1 v1.2 versus Amplicor HIV-1 DNA Test v1.5) for Early Diagnosis of HIV-1 Infection in Infants in Senegal

“…Several techniques for detecting proviral DNA use DBS for early infant HIV-1 diagnosis (11,13,15,23,29). Some have been developed in-house, while others are commercial, such as the validated Amplicor technique, which has sensitivity on DBS of 98.2 to 100% and a specificity of 98.6 to 100% (15,16,23). The high sensitivity of plasma HIV RNA quantification techniques now allows their substitution for DNA PCR tests (10,22,28).…”

“…DBS are easy to obtain and are a convenient method for collecting and transporting blood samples because they have less restrictive transport requirements than whole blood in tubes. Several techniques for detecting proviral DNA use DBS for early infant HIV-1 diagnosis (11,13,15,23,29). Some have been developed in-house, while others are commercial, such as the validated Amplicor technique, which has sensitivity on DBS of 98.2 to 100% and a specificity of 98.6 to 100% (15,16,23).…”