AIDSWEEKLY Plus; Monday, June 27, 2005
Staff Medical Writers

According to published research from the United States, "A dried blood spot (DBS) is a well-accepted means for the collection, transport, and storage of blood samples for various epidemiologic, serologic, and molecular assays for human immunodeficiency virus (HIV) studies. It is particularly important for mother-to-infant-transmission studies of affected individuals living in remote areas."

"We have developed a real-time PCR method to detect HIV type 1 (HIV-1) DNA in dried blood spots. A cellular gene, RNase P, was coamplified with the HIV-1 DNA in the same tube to monitor the DNA extraction efficiency and the overall assay performance.

"Our assay is a one-tube, single-step closed-system assay and uses a dUTP/uracil DNA glycosidase anti-PCR contamination control. The HIV-1 primers and probe were derived from a conserved region of the long terminal repeat," wrote W. Luo and colleagues at the U.S. Centers for Disease Control and Prevention in Atlanta.

"The detection of RNase P is attenuated by lowering the forward and reverse primer concentrations so that its amplification will not overwhelm the HIV-1 amplification and yet will provide a semiquantitative measurement of the quality of the isolated DBS DNA," scientists said.

"We examined 103 HIV-1-seropositive and 56 seronegative U.S. adults and found that our assay has a sensitivity of 98.1% (95% confidence interval [CI], 95.5 to 100%) and specificity of 100% (95% CI, 99 to 100%). The positive and negative predictive values are 100 and 96.6%, respectively," reported the authors.

Luo concluded, "This duplex PCR assay may be useful in identifying HIV-1-infected persons, particularly infants born to seropositive mothers in remote areas of the world."

Luo and colleagues published their findings in the Journal of Clinical Microbiology (Detection of human immunodeficiency virus type 1 DNA in dried blood spots by a duplex real-time PCR assay. J Clin Microbiol. 2005 Apr;43(4):1851-7.

Additional information can be obtained by contacting C.Y. Ou, Mailstop A-25, 16000 Clifton Rd., Atlanta, GA 30333, USA.

The publisher of the Journal of Clinical Microbiology can be contacted at: American Society Microbiology, 1752 N St. NW, Washington, DC 20036-2904, USA.

Keywords: Atlanta, Georgia, United States, HIV/AIDS, Diagnostics, Viral Load, Dried Blood Spots, Real-Time PCR.

This article was prepared by AIDS Weekly editors from staff and other reports.

Reference

Luo W, Yang H, Rathbun K, et al., Detection of human immunodeficiency virus type 1 DNA in dried blood spots by a duplex real-time PCR assay, J Clin Microbiol. 2005 Apr;43(4):1851-7.

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