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Published on: July 9, 2014
Identifying human immunodeficiency virus infection at birth: application of polymerase chain reaction to Guthrie
A M Comeau1, H W Hsu, M Schwerzler
1Newborn Screening Division, State Laboratory Institute of Massachusetts, Jamaica Plain 02130.
Insights
Polymerase chain reaction (PCR) detected human immunodeficiency virus (HIV) proviral DNA in 52% of infected newborns using dried blood spots. This method shows promise for early HIV diagnosis and epidemiological studies in infants.
Area of Science:
- Virology
- Pediatric Infectious Diseases
- Molecular Diagnostics
Background:
- Early diagnosis of pediatric HIV infection is crucial for timely intervention and improved outcomes.
- Traditional HIV antibody tests may not be reliable in newborns due to maternal antibodies.
- Dried blood spots (Guthrie cards) are a valuable resource for retrospective neonatal screening.
Purpose of the Study:
- To evaluate the utility of the polymerase chain reaction (PCR) assay for detecting HIV proviral DNA in dried blood spots from neonates.
- To determine if PCR can predict HIV infection at birth.
- To assess the correlation between PCR positivity and clinical progression in infected children.
Main Methods:
- Retrospective analysis of archived Guthrie cards from 1986-1991.
- Samples included 67 HIV-infected children and 63 children with seroreversion.
- High-sensitivity PCR assay (detecting 2-10 copies of HIV proviral DNA/µg) was employed.
Main Results:
- HIV proviral DNA was detected in 52% (35/67) of infected neonatal specimens.
- Longer storage times did not negatively impact PCR positivity rates.
- PCR positivity was higher in neonates who later showed aggressive clinical progression, including low CD4 counts, Pneumocystis pneumonia, or acquired immunodeficiency syndrome (AIDS).
Conclusions:
- PCR analysis of neonatal dried blood spots is a viable method for early HIV diagnosis.
- The technique demonstrates potential for epidemiological projections beyond seroprevalence studies.
- PCR offers an advantage over antibody assays due to its ability to detect viral DNA directly.
Abstract:
Guthrie cards containing dried blood spots from 67 children now known to be infected with human immunodeficiency virus (HIV) and 63 children now classified as having had seroreversion were retrieved from the newborn infant archives from 1986 through 1991 to determine whether the polymerase chain reaction (PCR) could predict the infection at birth. The PCR assays operating at a sensitivity capable of detecting 2 to 10 copies of HIV proviral DNA per microgram were able to detect HIV proviral DNA in 52% (35/67) of the infected neonatal blood specimens. Longer storage times did not decrease PCR positivity rates, an advantage over assays for HIV antibody. Children whose clinical progression has been aggressive had high rates of PCR positivity in neonatal specimens, 50% (7/14) in those with low CD4 cell counts during the first year of life, 71% (10/14) in those with Pneumocystis pneumonia or disseminated cytomegalovirus infection by age 1 year, 62% (18/29) in those with onset of acquired immunodeficiency syndrome by 18 months, and 66% (14/21) in those who died of the disease by 36 months of age. No evidence of HIV proviral DNA was found in any of the 63 specimens from children with seroreversion. We conclude that PCR, using routinely available dried blood spots from neonates, has applications in early diagnosis and in epidemiologic projections going beyond current seroprevalence studies.
