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Enzymatic amplification of the human immunodeficiency virus in peripheral blood mononuclear cells from pediatric
E G Chadwick1, R Yogev, S Kwok
1Department of Pediatrics, Northwestern University Medical School, Chicago, Illinois.
Insights
Polymerase chain reaction (PCR) can detect human immunodeficiency virus type 1 (HIV) provirus in infants. This method accurately identifies HIV infection in newborns and children, distinguishing them from uninfected infants.
Area of Science:
- Virology
- Immunology
- Pediatrics
Background:
- Vertical transmission of human immunodeficiency virus type 1 (HIV) poses a significant risk to infants born to infected mothers.
- Accurate and early diagnosis of HIV infection in infants is crucial for timely intervention and management.
Purpose of the Study:
- To evaluate the utility of the polymerase chain reaction (PCR) technique for detecting HIV-1 provirus in peripheral blood mononuclear cells (PBMCs) of infants.
- To assess the diagnostic performance of PCR in differentiating infected from uninfected infants born to HIV-positive mothers.
Main Methods:
- Peripheral blood mononuclear cells (PBMCs) were collected from HIV-infected mothers, their offspring, and control mother-infant pairs.
- HIV-1 proviral DNA was amplified from PBMC lysates using PCR targeting conserved gag and env gene regions.
- Amplified products were detected using an oligomer hybridization procedure.
Main Results:
- HIV-1 proviral DNA was detected in PBMCs of infants confirmed as infected by Centers for Disease Control criteria and in their HIV-infected mothers.
- Clinically uninfected infants who were seropositive with declining antibody titers or became seronegative lacked detectable HIV-1 proviral DNA.
- PCR demonstrated a characteristic HIV oligomer hybridization product in infected individuals.
Conclusions:
- Polymerase chain reaction (PCR) is a valuable diagnostic tool for detecting or excluding HIV-1 infection in young infants and children.
- PCR offers a reliable method for early diagnosis of HIV infection in newborns, complementing traditional serological methods.
- The presence or absence of amplified HIV-1 gene products correlates with infection status in infants.
Abstract:
The presence of the human immunodeficiency virus type 1 (HIV) provirus was assessed in peripheral blood mononuclear cells (PBMCs) from 27 offspring of HIV-infected mothers, 12 of these mothers, and 4 HIV-uninfected mother-infant control pairs. Enzymatic amplification of specific conserved regions of the gag and env genes was performed directly in PBMC lysates using the polymerase chain reaction (PCR) technique. The enzymatically amplified gene products were evaluated using the oligomer hybridization detection procedure. PBMCs from infected infants (as determined by Centers for Disease Control clinical criteria) and from HIV-infected mothers manifested a characteristic HIV oligomer hybridization product. Clinically uninfected seropositive infants with declining HIV antibody titers and infants who became seronegative lacked an enzymatically amplified HIV gene product. These preliminary data indicate that PCR is a valuable diagnostic technique to detect or exclude HIV infection in young infants and children.