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Low-cost HIV-1 diagnosis and quantification in dried blood spots by real time PCR. | LitMetric

AI Article Synopsis

  • Rapid and cost-effective HIV-1 diagnostic methods are essential for managing infections in low-resource settings, especially for early treatment recommendations for perinatally infected infants.
  • A new real-time LightCycler PCR assay has been developed to detect and quantify HIV-1 using dried blood spots (DBS), showing excellent sensitivity and correlation with standard commercial methods.
  • This rtLC DBS assay is significantly faster and cheaper than existing assays, making it a promising tool for early diagnosis and monitoring of HIV-1, particularly in pediatric cases in underserved areas.

Article Abstract

Background: Rapid and cost-effective methods for HIV-1 diagnosis and viral load monitoring would greatly enhance the clinical management of HIV-1 infected adults and children in limited-resource settings. Recent recommendations to treat perinatally infected infants within the first year of life are feasible only if early diagnosis is routinely available. Dried blood spots (DBS) on filter paper are an easy and convenient way to collect and transport blood samples. A rapid and cost effective method to diagnose and quantify HIV-1 from DBS is urgently needed to facilitate early diagnosis of HIV-1 infection and monitoring of antiretroviral therapy.

Methods And Findings: We have developed a real-time LightCycler (rtLC) PCR assay to detect and quantify HIV-1 from DBS. HIV-1 RNA extracted from DBS was amplified in a one-step, single-tube system using primers specific for long-terminal repeat sequences that are conserved across all HIV-1 clades. SYBR Green dye was used to quantify PCR amplicons and HIV-1 RNA copy numbers were determined from a standard curve generated using serially diluted known copies of HIV-1 RNA. This assay detected samples across clades, has a dynamic range of 5 log(10), and %CV <8% up to 4 log(10) dilution. Plasma HIV-1 RNA copy numbers obtained using this method correlated well with the Roche Ultrasensitive (r = 0.91) and branched DNA (r = 0.89) assays. The lower limit of detection (95%) was estimated to be 136 copies. The rtLC DBS assay was 2.5 fold rapid as well as 40-fold cheaper when compared to commercial assays. Adaptation of the assay into other real-time systems demonstrated similar performance.

Conclusions: The accuracy, reliability, genotype inclusivity and affordability, along with the small volumes of blood required for the assay suggest that the rtLC DBS assay will be useful for early diagnosis and monitoring of pediatric HIV-1 infection in resource-limited settings.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2688035PMC
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0005819PLOS

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