Detection of HIV-1 sequences in children using radioactive and colorimetric polymerase chain reactions

Area of Science:

• Virology
• Molecular Biology
• Pediatric Infectious Diseases

Background:

• Early diagnosis of Human Immunodeficiency Virus type 1 (HIV-1) infection in children is crucial for timely intervention and improved outcomes.
• Vertical transmission from mother to child remains a significant route of pediatric HIV-1 infection.

Purpose of the Study:

• To evaluate the efficacy of polymerase chain reaction (PCR) for detecting HIV-1 proviral DNA in infants born to HIV-1-seropositive mothers.
• To compare a novel colorimetric microwell detection method with a traditional radioactive electrophoretic method for PCR-based HIV-1 diagnosis.

Main Methods:

• Peripheral blood mononuclear cells from 18 HIV-1 infected and 28 uninfected children were analyzed using PCR to detect a gag region DNA fragment.
• PCR products were detected using either a radioactive probe or a tagged probe for colorimetric microwell visualization.

Main Results:

• The radioactive PCR method showed 100% specificity and was more sensitive than the p24 antigen test, though two false negatives occurred.
• The colorimetric detection method demonstrated 100% specificity and comparable sensitivity to the radioactive method, with four discordant results in limited comparative analysis.
• PCR proved superior for early diagnosis of pediatric HIV-1 infection, with the colorimetric assay showing potential for routine use.

Conclusions:

• Polymerase chain reaction (PCR) is the optimal method for the early diagnosis of HIV-1 infection in pediatric populations.
• The developed colorimetric microwell plate procedure is a valuable and potentially routine diagnostic technique for pediatric HIV-1 detection.