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. 1989 Jan;27(1):120–125. doi: 10.1128/jcm.27.1.120-125.1989

Monoclonal antibody solution hybridization assay for detection of human immunodeficiency virus nucleic acids.

R P Viscidi 1, C O'Meara 1, H Farzadegan 1, R Yolken 1
PMCID: PMC267246  PMID: 2913023

Abstract

In this report we describe a novel, nonisotopic hybridization assay for the measurement of viral RNA in biological samples. The assay involved a solution-phase reaction between a biotinylated DNA probe and RNA target sequences. Labeled hybrids were detected in an immunoreaction by using a solid-phase anti-biotin antibody and an enzyme-labeled monoclonal antibody specific for DNA-RNA hybrids. This monoclonal antibody solution hybridization assay was compared with an antigen-capture immunoassay for the detection of human immunodeficiency virus in 436 cell culture samples from 60 seropositive patients. The sensitivity and specificity of the hybridization assay were 93.5 and 94.6%, respectively. Detection of human immunodeficiency virus solely by hybridization in the initial sample but not subsequent samples from seven cultures may reflect detection of virus that was present in the patients' lymphocytes but did not replicate in vitro. Since the assay method is adapatable to the detection of either RNA or DNA, it could provide a means for the detection of a wide range of viral nucleic acids.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boguslawski S. J., Smith D. E., Michalak M. A., Mickelson K. E., Yehle C. O., Patterson W. L., Carrico R. J. Characterization of monoclonal antibody to DNA.RNA and its application to immunodetection of hybrids. J Immunol Methods. 1986 May 1;89(1):123–130. doi: 10.1016/0022-1759(86)90040-2. [DOI] [PubMed] [Google Scholar]
  2. Chollet A., Kawashima E. H. Biotin-labeled synthetic oligodeoxyribonucleotides: chemical synthesis and uses as hybridization probes. Nucleic Acids Res. 1985 Mar 11;13(5):1529–1541. doi: 10.1093/nar/13.5.1529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Forster A. C., McInnes J. L., Skingle D. C., Symons R. H. Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotin. Nucleic Acids Res. 1985 Feb 11;13(3):745–761. doi: 10.1093/nar/13.3.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gallo R. C., Salahuddin S. Z., Popovic M., Shearer G. M., Kaplan M., Haynes B. F., Palker T. J., Redfield R., Oleske J., Safai B. Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Science. 1984 May 4;224(4648):500–503. doi: 10.1126/science.6200936. [DOI] [PubMed] [Google Scholar]
  5. Goudsmit J., de Wolf F., Paul D. A., Epstein L. G., Lange J. M., Krone W. J., Speelman H., Wolters E. C., Van der Noordaa J., Oleske J. M. Expression of human immunodeficiency virus antigen (HIV-Ag) in serum and cerebrospinal fluid during acute and chronic infection. Lancet. 1986 Jul 26;2(8500):177–180. doi: 10.1016/s0140-6736(86)92485-2. [DOI] [PubMed] [Google Scholar]
  6. Kempe T., Sundquist W. I., Chow F., Hu S. L. Chemical and enzymatic biotin-labeling of oligodeoxyribonucleotides. Nucleic Acids Res. 1985 Jan 11;13(1):45–57. doi: 10.1093/nar/13.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Levy J. A., Hoffman A. D., Kramer S. M., Landis J. A., Shimabukuro J. M., Oshiro L. S. Isolation of lymphocytopathic retroviruses from San Francisco patients with AIDS. Science. 1984 Aug 24;225(4664):840–842. doi: 10.1126/science.6206563. [DOI] [PubMed] [Google Scholar]
  8. Levy J. A., Shimabukuro J. Recovery of AIDS-associated retroviruses from patients with AIDS or AIDS-related conditions and from clinically healthy individuals. J Infect Dis. 1985 Oct;152(4):734–738. doi: 10.1093/infdis/152.4.734. [DOI] [PubMed] [Google Scholar]
  9. Markham P. D., Salahuddin S. Z., Popovic M., Patel A., Veren K., Fladager A., Orndorff S., Gallo R. C. Advances in the isolation of HTLV-III from patients with AIDS and AIDS-related complex and from donors at risk. Cancer Res. 1985 Sep;45(9 Suppl):4588s–4591s. [PubMed] [Google Scholar]
  10. Renz M., Kurz C. A colorimetric method for DNA hybridization. Nucleic Acids Res. 1984 Apr 25;12(8):3435–3444. doi: 10.1093/nar/12.8.3435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Richman D. D., McCutchan J. A., Spector S. A. Detecting human immunodeficiency virus RNA in peripheral blood mononuclear cells by nucleic acid hybridization. J Infect Dis. 1987 Nov;156(5):823–827. doi: 10.1093/infdis/156.5.823. [DOI] [PubMed] [Google Scholar]
  12. Tchen P., Fuchs R. P., Sage E., Leng M. Chemically modified nucleic acids as immunodetectable probes in hybridization experiments. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3466–3470. doi: 10.1073/pnas.81.11.3466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Viscidi R. P., Connelly C. J., Yolken R. H. Novel chemical method for the preparation of nucleic acids for nonisotopic hybridization. J Clin Microbiol. 1986 Feb;23(2):311–317. doi: 10.1128/jcm.23.2.311-317.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Viscidi R. P., Yolken R. G. Molecular diagnosis of infectious diseases by nucleic acid hybridization. Mol Cell Probes. 1987 Mar;1(1):3–14. doi: 10.1016/0890-8508(87)90003-x. [DOI] [PubMed] [Google Scholar]
  15. Viscidi R., Farzadegan H., Leister F., Francisco M. L., Yolken R. Enzyme immunoassay for detection of human immunodeficiency virus antigens in cell cultures. J Clin Microbiol. 1988 Mar;26(3):453–458. doi: 10.1128/jcm.26.3.453-458.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yehle C. O., Patterson W. L., Boguslawski S. J., Albarella J. P., Yip K. F., Carrico R. J. A solution hybridization assay for ribosomal RNA from bacteria using biotinylated DNA probes and enzyme-labeled antibody to DNA:RNA. Mol Cell Probes. 1987 Jun;1(2):177–193. doi: 10.1016/0890-8508(87)90026-0. [DOI] [PubMed] [Google Scholar]

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