Transmission of HIV-1 in infants born to seropositive mothers: PCR-amplified proviral DNA detected by flow cytometric analysis of immunoreactive beads.

The diagnosis of HIV infection in newborns is established by amplification of proviral DNA using the polymerase chain reaction (PCR). We developed a nonisotopic method for heminested PCR using a biotinylated primer among sets of three oligonucleotides, each selected from the HIV long terminal repeat (LTR) and gag sequences. An internal probe incorporating digoxigenin-dUTP was also synthesized by PCR. The PCR products, hybridized with LTR region or gag region probes, were captured with streptavidin-coated magnetic beads and detected by fluorescein isothiocyanate-labeled antidigoxigenin in flow cytometric analysis. This immunoreactive bead assay (PCR-IRB) detected about three copies of HIV proviral DNA. A panel of 50 coded DNA specimens of infants previously assayed by conventional PCR and with known clinical results revealed that the PCR-IRB findings using LTR, but not gag, were in agreement. A double-blind prospective study of blood samples from 14 mother-infant pairs using the PCR-IRB amplification of LTR gave results similar to the commercial Amplicor HIV-1 PCR test and were consistent with the clinical outcomes. PCR-IRB results were positive for 11 mothers and three infants, one at birth, one at 2 weeks after birth, and one at 8 weeks after birth. PCR-IRB is a simple, reliable, specific, and automatable assay useful in the early diagnosis of perinatal HIV infection in clinical practice and regional screening programs.

[1]  J. Bremer,et al.  Early detection of human immunodeficiency virus on dried blood spot specimens: sensitivity across serial specimens. Women and Infants Transmission Study Group. , 1996, The Journal of pediatrics.

[2]  H. Barnhart,et al.  Natural history of human immunodeficiency virus disease in perinatally infected children: an analysis from the Pediatric Spectrum of Disease Project. , 1996, Pediatrics.

[3]  B. Masquelier,et al.  Neonatal characteristics in rapidly progressive perinatally acquired HIV-1 disease , 1996 .

[4]  S. Pelton,et al.  Seroreversion in human immunodeficiency virus‐exposed but uninfected infants , 1995, The Pediatric infectious disease journal.

[5]  D. Gibb,et al.  A Risk-Benefit Assessment of Zidovudine in the Prevention of Perinatal HIV Transmission , 1995, Drug safety.

[6]  R. Gelber,et al.  Reduction of maternal-infant transmission of human immunodeficiency virus type 1 with zidovudine treatment. Pediatric AIDS Clinical Trials Group Protocol 076 Study Group. , 1994, The New England journal of medicine.

[7]  Chrismon Jh,et al.  Surveillance for smoking-attributable mortality and years of potential life lost, by state--United States, 1990. , 1994, MMWR. CDC surveillance summaries : Morbidity and mortality weekly report. CDC surveillance summaries.

[8]  G. Vyas,et al.  Flow cytometric immunodetection of human immunodeficiency virus type 1 proviral DNA by heminested PCR and digoxigenin-labeled probes , 1994, Clinical and diagnostic laboratory immunology.

[9]  E. Walter,et al.  Identifying human immunodeficiency virus infection at birth: application of polymerase chain reaction to Guthrie cards. , 1993, The Journal of pediatrics.

[10]  J. Church.  Clinical aspects of HIV infection in children. , 1993, Pediatric annals.

[11]  R. Aiyer,et al.  Detection of hepatitis B virus in plasma using flow cytometric analyses of polymerase chain reaction-amplified DNA incorporating digoxigenin-11-dUTP. , 1993, Blood.

[12]  E. R. Stiehm,et al.  Rapid serologic testing with immune‐complex‐dissociated HIV p24 antigen for early detection of HIV infection in neonates , 1993, The New England journal of medicine.

[13]  G. Vyas,et al.  An improved method for the detection of hepatitis C virus RNA in plasma utilizing heminested primers and internal control RNA. , 1993, PCR methods and applications.

[14]  W. Shearer,et al.  Sensitivity, specificity and predictive value of physical examination, culture and other laboratory studies in the diagnosis during early infancy of vertically acquired human immunodeficiency virus infection , 1993, The Pediatric infectious disease journal.

[15]  G. Schutze,et al.  Routine tuberculin screening of children during hospitalization , 1993, The Pediatric infectious disease journal.

[16]  D. Wara,et al.  Proposed definitions for in utero versus intrapartum transmission of HIV-1. , 1992, The New England journal of medicine.

[17]  W. Borkowsky,et al.  Early diagnosis of human immunodeficiency virus infection in children less than 6 months of age: comparison of polymerase chain reaction, culture, and plasma antigen capture techniques. , 1992, The Journal of infectious diseases.

[18]  D. Bylund,et al.  Review of testing for human immunodeficiency virus. , 1992, Clinics in laboratory medicine.

[19]  J. Levy,et al.  Polymerase chain reaction compared with concurrent viral cultures for rapid identification of human immunodeficiency virus infection among high-risk infants and children. , 1989, The Journal of pediatrics.

[20]  P. Selwyn,et al.  Use of the polymerase chain reaction for early detection of the proviral sequences of human immunodeficiency virus in infants born to seropositive mothers , 1989, Disease markers.

[21]  A. Fauci,et al.  Monokine regulation of human immunodeficiency virus-1 expression in a chronically infected human T cell clone. , 1989, Journal of immunology.