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. 1985 Sep-Oct;58(5):425–442.

Principles of nucleic acid hybridization and comparison with monoclonal antibody technology for the diagnosis of infectious diseases.

S C Edberg
PMCID: PMC2589955  PMID: 3004048

Abstract

Until the 1980s the diagnosis of specific etiologic agents of infectious diseases rested with their isolation in vitro and identification by analysis of their phenotypic characteristics. In the 1970s the concept of a microbial species evolved from phenotypic analysis to nucleic acid homology. Currently, nucleic acid sequences specific for a given species are being isolated and amplified and utilized not only to identify the pathogen after it has been grown in vitro but also elucidate it directly in biological material. The procedures for making nucleic acid hybridization probes are analogous to the generation of monoclonal antibody tests. Currently, research and development are centered in choosing the particular nucleic acid to analyze, establishing the most efficient vector system for amplifying the nucleic acid, generating an efficient means of selecting the particular nucleic acid fragment specific for the microorganism, and in measuring the hybridization reaction. While immunological techniques have been utilized in the clinical laboratory for over thirty years, the means of detecting nucleic acid hybridization reactions are just beginning to be usable in the clinical diagnostic laboratory. Much of nucleic acid hybridization research is proprietary, and a particular challenge is to develop a means whereby information can be used for the progress of science as a whole when generated by private ownership.

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Selected References

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

  1. Andiman W., Gradoville L., Heston L., Neydorff R., Savage M. E., Kitchingman G., Shedd D., Miller G. Use of cloned probes to detect Epstein-Barr viral DNA in tissues of patients with neoplastic and lymphoproliferative diseases. J Infect Dis. 1983 Dec;148(6):967–977. doi: 10.1093/infdis/148.6.967. [DOI] [PubMed] [Google Scholar]
  2. Bauman J. G., Wiegant J., van Duijn P. Cytochemical hybridisation with fluorochrome-labelled RNA. III. Increased sensitivity by the use of anti-fluorescein antibodies. Histochemistry. 1981;73(2):181–193. doi: 10.1007/BF00493018. [DOI] [PubMed] [Google Scholar]
  3. Bauman J. G., Wiegant J., van Duijn P. Cytochemical hybridization with fluorochrome-labeled RNA. II. Applications. J Histochem Cytochem. 1981 Feb;29(2):238–246. doi: 10.1177/29.2.6166654. [DOI] [PubMed] [Google Scholar]
  4. Bayer E. A., Wilchek M. The use of the avidin-biotin complex as a tool in molecular biology. Methods Biochem Anal. 1980;26:1–45. doi: 10.1002/9780470110461.ch1. [DOI] [PubMed] [Google Scholar]
  5. Berninger M., Hammer M., Hoyer B., Gerin J. L. An assay for the detection of the DNA genome of hepatitis B virus in serum. J Med Virol. 1982;9(1):57–68. doi: 10.1002/jmv.1890090109. [DOI] [PubMed] [Google Scholar]
  6. Brandsma J., Miller G. Nucleic acid spot hybridization: rapid quantitative screening of lymphoid cell lines for Epstein-Barr viral DNA. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6851–6855. doi: 10.1073/pnas.77.11.6851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brechot C., Hadchouel M., Scotto J., Degos F., Charnay P., Trepo C., Tiollais P. Detection of hepatitis B virus DNA in liver and serum: a direct appraisal of the chronic carrier state. Lancet. 1981 Oct 10;2(8250):765–768. doi: 10.1016/s0140-6736(81)90182-3. [DOI] [PubMed] [Google Scholar]
  8. Bresser J., Doering J., Gillespie D. Quick-blot: selective mRNA or DNA immobilization from whole cells. DNA. 1983;2(3):243–254. doi: 10.1089/dna.1983.2.243. [DOI] [PubMed] [Google Scholar]
  9. Brigati D. J., Myerson D., Leary J. J., Spalholz B., Travis S. Z., Fong C. K., Hsiung G. D., Ward D. C. Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probes. Virology. 1983 Apr 15;126(1):32–50. doi: 10.1016/0042-6822(83)90460-9. [DOI] [PubMed] [Google Scholar]
  10. Casna N. J., Shub D. A. Bacteriophage T4 as a generalized DNA-cloning vehicle. Gene. 1982 Jun;18(3):297–307. doi: 10.1016/0378-1119(82)90168-8. [DOI] [PubMed] [Google Scholar]
  11. Cederbaum S. D. Introduction to recombinant DNA. Pediatrics. 1984 Sep;74(3):408–411. [PubMed] [Google Scholar]
  12. Chou S., Merigan T. C. Rapid detection and quantitation of human cytomegalovirus in urine through DNA hybridization. N Engl J Med. 1983 Apr 21;308(16):921–925. doi: 10.1056/NEJM198304213081603. [DOI] [PubMed] [Google Scholar]
  13. Coonrod J. D. Urine as an antigen reservoir for diagnosis of infectious diseases. Am J Med. 1983 Jul 28;75(1B):85–92. doi: 10.1016/0002-9343(83)90077-3. [DOI] [PubMed] [Google Scholar]
  14. Crosson F. J., Jr, Winkelstein J. A., Moxon E. R. Enzyme-linked immunosorbent assay for detection and quantitation of capsular antigen of Haemophilus influenzae type b. Infect Immun. 1978 Nov;22(2):617–619. doi: 10.1128/iai.22.2.617-619.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Drow D. L., Maki D. G., Manning D. D. Indirect sandwich enzyme-linked immunosorbent assay for rapid detection of Haemophilus influenzae type b infection. J Clin Microbiol. 1979 Oct;10(4):442–450. doi: 10.1128/jcm.10.4.442-450.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Engleberg N. C., Eisenstein B. I. The impact of new cloning techniques on the diagnosis and treatment of infectious diseases. N Engl J Med. 1984 Oct 4;311(14):892–901. doi: 10.1056/NEJM198410043111406. [DOI] [PubMed] [Google Scholar]
  17. Goldstein L. C., Corey L., McDougall J. K., Tolentino E., Nowinski R. C. Monoclonal antibodies to herpes simplex viruses: use in antigenic typing and rapid diagnosis. J Infect Dis. 1983 May;147(5):829–837. doi: 10.1093/infdis/147.5.829. [DOI] [PubMed] [Google Scholar]
  18. Green M. R., Maniatis T., Melton D. A. Human beta-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei. Cell. 1983 Mar;32(3):681–694. doi: 10.1016/0092-8674(83)90054-5. [DOI] [PubMed] [Google Scholar]
  19. Grimont P. A., Grimont F., Desplaces N., Tchen P. DNA probe specific for Legionella pneumophila. J Clin Microbiol. 1985 Mar;21(3):431–437. doi: 10.1128/jcm.21.3.431-437.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hansen E. J., Robertson S. M., Gulig P. A., Frisch C. F., Haanes E. J. Immunoprotection of rats against Haemophilus influenzae type B disease mediated by monoclonal antibody against a haemophilus outer-membrane protein. Lancet. 1982 Feb 13;1(8268):366–368. doi: 10.1016/s0140-6736(82)91394-0. [DOI] [PubMed] [Google Scholar]
  21. Harding S. A., Scheld W. M., McGowan M. D., Sande M. A. Enzyme-linked immunosorbent assay for detection of Streptococcus pneumoniae antigen. J Clin Microbiol. 1979 Sep;10(3):339–342. doi: 10.1128/jcm.10.3.339-342.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hill W. E., Madden J. M., McCardell B. A., Shah D. B., Jagow J. A., Payne W. L., Boutin B. K. Foodborne enterotoxigenic Escherichia coli: detection and enumeration by DNA colony hybridization. Appl Environ Microbiol. 1983 Apr;45(4):1324–1330. doi: 10.1128/aem.45.4.1324-1330.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  24. Kaplan S. L. Antigen detection in cerebrospinal fluid--pros and cons. Am J Med. 1983 Jul 28;75(1B):109–118. doi: 10.1016/0002-9343(83)90081-5. [DOI] [PubMed] [Google Scholar]
  25. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  26. Langer P. R., Waldrop A. A., Ward D. C. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633–6637. doi: 10.1073/pnas.78.11.6633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Leary J. J., Brigati D. J., Ward D. C. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045–4049. doi: 10.1073/pnas.80.13.4045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lefèvre J. F., Fuchs R. P., Daune M. P. Comparative studies on the 7-iodo and 7-fluoro derivatives of N-acetoxy-N-2-acetylaminofluorene: binding sites on DNA and conformational change of modified deoxytrinucleotides. Biochemistry. 1978 Jun 27;17(13):2561–2567. doi: 10.1021/bi00606a016. [DOI] [PubMed] [Google Scholar]
  29. MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J Mol Biol. 1962 Jul;5:109–118. doi: 10.1016/s0022-2836(62)80066-7. [DOI] [PubMed] [Google Scholar]
  30. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Manuelidis L., Langer-Safer P. R., Ward D. C. High-resolution mapping of satellite DNA using biotin-labeled DNA probes. J Cell Biol. 1982 Nov;95(2 Pt 1):619–625. doi: 10.1083/jcb.95.2.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Moseley S. L., Echeverria P., Seriwatana J., Tirapat C., Chaicumpa W., Sakuldaipeara T., Falkow S. Identification of enterotoxigenic Escherichia coli by colony hybridization using three enterotoxin gene probes. J Infect Dis. 1982 Jun;145(6):863–869. doi: 10.1093/infdis/145.6.863. [DOI] [PubMed] [Google Scholar]
  33. Moseley S. L., Huq I., Alim A. R., So M., Samadpour-Motalebi M., Falkow S. Detection of enterotoxigenic Escherichia coli by DNA colony hybridization. J Infect Dis. 1980 Dec;142(6):892–898. doi: 10.1093/infdis/142.6.892. [DOI] [PubMed] [Google Scholar]
  34. Myerson D., Hackman R. C., Meyers J. D. Diagnosis of cytomegaloviral pneumonia by in situ hybridization. J Infect Dis. 1984 Aug;150(2):272–277. doi: 10.1093/infdis/150.2.272. [DOI] [PubMed] [Google Scholar]
  35. Pepple J., Moxon E. R., Yolken R. H. Indirect enzyme-linked immunosorbent assay for the quantitation of the type-specific antigen of Haemophilus influenzae b: a preliminary report. J Pediatr. 1980 Aug;97(2):233–237. doi: 10.1016/s0022-3476(80)80480-x. [DOI] [PubMed] [Google Scholar]
  36. Polin R. A., Kennett R. Use of monoclonal antibodies in an enzyme-linked inhibition assay for rapid detection of streptococcal antigen. J Pediatr. 1980 Oct;97(4):540–544. doi: 10.1016/s0022-3476(80)80005-9. [DOI] [PubMed] [Google Scholar]
  37. Porterfield J. S., Tobin J. O. Viral and bacterial infectious diseases. Br Med Bull. 1984 Jul;40(3):283–290. doi: 10.1093/oxfordjournals.bmb.a071990. [DOI] [PubMed] [Google Scholar]
  38. Ranki M., Palva A., Virtanen M., Laaksonen M., Söderlund H. Sandwich hybridization as a convenient method for the detection of nucleic acids in crude samples. Gene. 1983 Jan-Feb;21(1-2):77–85. doi: 10.1016/0378-1119(83)90149-x. [DOI] [PubMed] [Google Scholar]
  39. Redfield D. C., Richman D. D., Albanil S., Oxman M. N., Wahl G. M. Detection of herpes simplex virus in clinical specimens by DNA hybridization. Diagn Microbiol Infect Dis. 1983 Jun;1(2):117–128. doi: 10.1016/0732-8893(83)90041-x. [DOI] [PubMed] [Google Scholar]
  40. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  41. Schildkraut C. Dependence of the melting temperature of DNA on salt concentration. Biopolymers. 1965;3(2):195–208. doi: 10.1002/bip.360030207. [DOI] [PubMed] [Google Scholar]
  42. Scotto J., Hadchouel M., Hery C., Yvart J., Tiollais P., Brechot C. Detection of hepatitis B virus DNA in serum by a simple spot hybridization technique: comparison with results for other viral markers. Hepatology. 1983 May-Jun;3(3):279–284. doi: 10.1002/hep.1840030301. [DOI] [PubMed] [Google Scholar]
  43. Sippel J. E., Voller A. Detection of Neisseria meningitidis cell envelope antigen by enzyme-linked immunosorbent assay in patients with meningococcal disease. Trans R Soc Trop Med Hyg. 1980;74(5):644–648. doi: 10.1016/0035-9203(80)90156-x. [DOI] [PubMed] [Google Scholar]
  44. Spector S. A., Rua J. A., Spector D. H., McMillan R. Detection of human cytomegalovirus in clinical specimens by DNA-DNA hybridization. J Infect Dis. 1984 Jul;150(1):121–126. doi: 10.1093/infdis/150.1.121. [DOI] [PubMed] [Google Scholar]
  45. Stålhandske P., Hyypiä T., Gadler H., Halonen P., Pettersson U. The use of molecular hybridization for demonstration of adenoviruses in human stools. Curr Top Microbiol Immunol. 1983;104:299–306. doi: 10.1007/978-3-642-68949-9_19. [DOI] [PubMed] [Google Scholar]
  46. 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]
  47. Torres A. R., Li M. K., Ward D. C., Edberg S. C. Differentiation of Neisseria gonorrhoeae from other Neisseria species by use of the restriction endonuclease HaeIII. J Clin Microbiol. 1984 Oct;20(4):687–690. doi: 10.1128/jcm.20.4.687-690.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Totten P. A., Holmes K. K., Handsfield H. H., Knapp J. S., Perine P. L., Falkow S. DNA hybridization technique for the detection of Neisseria gonorrhoeae in men with urethritis. J Infect Dis. 1983 Sep;148(3):462–471. doi: 10.1093/infdis/148.3.462. [DOI] [PubMed] [Google Scholar]
  49. Virtanen M., Palva A., Laaksonen M., Halonen P., Söderlund H., Ranki M. Novel test for rapid viral diagnosis: detection of adenovirus in nasopharyngeal mucus aspirates by means of nucleic-acid sandwich hybridisation. Lancet. 1983 Feb 19;1(8321):381–383. doi: 10.1016/s0140-6736(83)91500-3. [DOI] [PubMed] [Google Scholar]
  50. Weller I. V., Fowler M. J., Monjardino J., Thomas H. C. The detection of HBV-DNA in serum by molecular hybridisation: a more sensitive method for the detection of complete HBV particles. J Med Virol. 1982;9(4):273–280. doi: 10.1002/jmv.1890090405. [DOI] [PubMed] [Google Scholar]
  51. Wetherall B. L., Hallsworth P. G., McDonald P. J. Enzyme-linked immunosorbent assay for detection of Haemophilus influenzae type b antigen. J Clin Microbiol. 1980 Jun;11(6):573–578. doi: 10.1128/jcm.11.6.573-578.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wetmur J. G., Davidson N. Kinetics of renaturation of DNA. J Mol Biol. 1968 Feb 14;31(3):349–370. doi: 10.1016/0022-2836(68)90414-2. [DOI] [PubMed] [Google Scholar]
  53. Yolken R. H. Enzyme immunoassays for the detection of infectious antigens in body fluids: current limitations and future prospects. Rev Infect Dis. 1982 Jan-Feb;4(1):35–68. doi: 10.1093/clinids/4.1.35. [DOI] [PubMed] [Google Scholar]
  54. Yolken R. H. Use of monoclonal antibodies for viral diagnosis. Curr Top Microbiol Immunol. 1983;104:177–195. doi: 10.1007/978-3-642-68949-9_11. [DOI] [PubMed] [Google Scholar]
  55. Zinn K., DiMaio D., Maniatis T. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene. Cell. 1983 Oct;34(3):865–879. doi: 10.1016/0092-8674(83)90544-5. [DOI] [PubMed] [Google Scholar]

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