Skip to main content
SpringerLink
Log in

Ultrasensitive Nonradioactive Detection of PCR Reactions: An Overview

  • Chapter
The Polymerase Chain Reaction

Abstract

Nonradioactive detection in one of its many forms has been the touchstone of modern nucleic acid diagnostics. Typically, these detection systems involve the determination of a chemical compound through the use of reactions that alter the characteristic light absorption or emission of a reporter molecule. Classical techniques that fall in this category include photometry, fluorescence, and chemi-luminescence, although one could argue that radioactivity might also be included since it is most often detected as a color change on X-ray film or as visible light in a liquid scintillation counter.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 59.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 79.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Arnold LJ, Hammond PW, Wiese WA, Nelson NC (1989): Assay formats involving acridinium ester labeled DNA probes. Clin Chem 35:1588–1594.

    CAS  PubMed  Google Scholar 

  • Balaguer P, Terouanne B, Eliaou JF, Humbert M, Boussioux AM, Nicolas JC (1989): Use of glu-cose-6-phosphate dehydrogenase as a new label for nucleic acid hybridization reactions. Anal Biochem 180(l):50–54.

    Article  CAS  PubMed  Google Scholar 

  • Balaguer P, Terouanne B, Boussioux AM, Nicolas JC (1989): Use of bioluminescence in nucleic acid hybridization reactions. J Biolumin Chemilumin 4(1):302–309.

    Article  CAS  PubMed  Google Scholar 

  • Ballagi-Pordany A, Klingeborn B, Flensburg J, Belak S (1990): Equine herpesvirus type 1: Detection of viral DNA sequences in aborted fetuses with the polymerase chain reaction. Vet Microbiol 22(4):373–381.

    Article  CAS  PubMed  Google Scholar 

  • Bauman JG, Wiegant J, van Duijn P (1983): The development, using poly (Hg-U) in a model system, of a new method to visualize hybridization in fluorescence microscopy. J Histochem Cytochem 31:571–578.

    Article  CAS  PubMed  Google Scholar 

  • Bej AK, Mahbubani MH, Miller R, DiCesare JL, Haff L, Atlas RM (1990): Multiplex PCR amplification and immobilized capture probes for detection of bacterial pathogens and indicators in water. Mol Cell Probes 4(5):353–365.

    Article  CAS  PubMed  Google Scholar 

  • Berninger M, Hammer M, Hoyer B, Gerin JL (1982): An assay for the detection of the DNA genome of hepatitis B virus in serum. J Med Virol 9:57–68.

    Article  CAS  PubMed  Google Scholar 

  • Brandsma J, Miller G (1980): Nucleic acid spot hybridization: Rapid quantitative screening of lymphoid cell lines for Epstein-Barr virus. Proc Natl Acad Sci USA 77:6851–6855.

    Article  CAS  PubMed  Google Scholar 

  • Brigati DJ, Myerson D, Leary JL, Spalholz B, Travis SZ, Fong CKY, Hsiung GD, Ward DC (1983): Detection of viral genomes in cultured cells and paraffin embeded tissue sections using biotin labeled hybridization probes. Virology 126:32–50.

    Article  CAS  PubMed  Google Scholar 

  • Brownlee RG, Sunzeri FJ, Busch MP (1990): Application of capillary DNA chromatography to detect AIDS virus (HIV-1) DNA in blood. J Chromatogr 533:87–96.

    Article  CAS  PubMed  Google Scholar 

  • Bryan RN, Arnold LJ Jr (1984): Detection of HBV-DNA with modified synthetic oligonucleotides containing radioactive and non-radioactive reporter groups. DNA 3:124.

    Google Scholar 

  • Bugawan TL, Erlich HA (1991): Rapid typing of HLA-DQB1 DNA polymorphism using nonradioactive oligonucleotide probes and amplified DNA. Immunogenetics 33(3): 163–170.

    Article  CAS  PubMed  Google Scholar 

  • Bugawan TL, Begovich AB, Erlich HA (1990): Rapid HLA-DPB typing using enzymatically amplified DNA and nonradioactive sequence-specific oligonucleotide probes. Immunogenetics 32(4):231–241.

    Article  CAS  PubMed  Google Scholar 

  • Chehab FF, Kan YW (1989): Detection of specific DNA sequences by fluorescence amplification: A color complementation assay. Proc Natl Acad Sci USA 86(23):9178–9182.

    Article  CAS  PubMed  Google Scholar 

  • Chehab FF, Kan YW (1990): Detection of sickle cell anaemia mutation by colour DNA amplification. Lancet 335(8680): 15–17.

    Article  CAS  PubMed  Google Scholar 

  • Chu BCF, Orgel LE (1985): Detection of specific DNA sequences with short biotin-labeled probes. DAW 4:327–331.

    CAS  Google Scholar 

  • Connolly BA, Rider P (1985): Chemical synthesis of oligonucleotides containing a free sulphydryl group and subsequent attachment of thiol specific probes. Nucl Acids Res 13:4485–4502.

    Article  CAS  PubMed  Google Scholar 

  • Coutlee F, Bobo L, Mayur K, Yolken RH, Viscidi RP (1989): Immunodetection of DNA with biotinylated RNA probes: A study of reactivity of a monoclonal antibody to DNA-RNA hybrids. Anal Biochem 181(1):96–105.

    Article  CAS  PubMed  Google Scholar 

  • Dahlen P, Iitia A, Mukkala VM, Hurskainen P, Kwiatkowski M (1991): The use of europium (Eu3 +) labelled primers in PCR amplification of specific target DNA. Mol Cell Probes 5(2): 143–149.

    Article  CAS  PubMed  Google Scholar 

  • Day PJ, Bevan IS, Gurney SJ, Young LS, Walker MR (1990): Synthesis in vitro and application of biotinylated DNA probes for human papilloma virus type 16 by utilizing the polymerase chain reaction. Biochem J 267(1): 119–123.

    CAS  PubMed  Google Scholar 

  • Dhingra K, Talpaz M, Riggs MG, Eastman PS, Zipf T, Ku S, Kurzrock R (1991): Hybridization protection assay: A rapid, sensitive, and specific method for detection of Philadelphia chromosome-positive leukemias. Blood 77(2):238–242.

    CAS  PubMed  Google Scholar 

  • Dissanayake S, Min X, Piessens WF (1991): Detection of amplified Wuchereria bancrofti DNA in mosquitoes with a nonradioactive probe. Mol Biochem Parasitol 45(1):49–56.

    Article  CAS  PubMed  Google Scholar 

  • Eliaou JF, Humbert M, Balaguer P, Gebuhrer L, Amsellem S, Betuel H, Nicolas JC, Clot J (1989): A method of HLA class II typing using nonradioactive labelled olignucleotides. Tissue Antigens 33(4):475–485.

    Article  CAS  PubMed  Google Scholar 

  • Embury SH, Kropp GL, Stanton TS, Warren TC, Cornett PA, Chehab FF (1990): Detection of the hemoglobin E mutation using the color complementation assay: Application to complex genotyping. Blood 76(3):619–623.

    CAS  PubMed  Google Scholar 

  • Erlich H, Bugawan T, Begovich AB, Scharf S, Griffith R, Saiki R, Higuchi R, Walsh PS (1991): HLA-DR, DQ and DP typing using PCR amplification and immobilized probes. Eur J Immunogen 18(1–2):33–55.

    Article  CAS  PubMed  Google Scholar 

  • Gardner L (1983): Non-radioactive DNA labeling: Detection of specific DNA and RNA sequences on nitrocellulose and in situ hybridizations. Bio Techniques 1:38–41.

    Article  CAS  Google Scholar 

  • Gregersen N, Winter V, Petersen KB, Koch J, Klvraa S, Rudiger N, Heinsvig EM, Bolund L (1989): Detection of point mutations in amplified single copy genes by biotin-labelled oligonucleotides: Diagnosis of variants of alpha-1-antitrypsin. Clin ChimActa 182(2): 151–164.

    Article  CAS  Google Scholar 

  • Heller MJ, Morrison LE (1985): Chemiluminescent and fluorescent probes for DNA hybridization systems. In: Rapid Detection and Identification of Infectious Agents, Kingsbury DT, Falkow S, eds. New York: Academic Press, 245–256.

    Google Scholar 

  • Heller MJ, Schneider BL (1983): Chemiluminescent labeling of nucleic acids using azidophenyl glyoxal. FedProc 42:1954 (abst. 1149).

    Google Scholar 

  • Hendy JG, Cauchi MN (1990): Direct detection of beta thalassemic mutations: Use of biotin labeled allele specific probes. Am J Hematol 34(2): 151–153.

    Article  CAS  PubMed  Google Scholar 

  • Hirschfeld T (1976): Optical microscope observation of single small molecules. Appl Optics 15:2965–2966.

    Article  CAS  Google Scholar 

  • Hsu SM, Raine L (1982): Versatility of biotin-labeled lectins and avidin-biotin-peroxidase complex for localization of carbohydrate in tissue sections. J Histochem Cytochem 30:157–161.

    Article  CAS  PubMed  Google Scholar 

  • Hsu SM, Raine L, Fanger H (1981): The use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques. A comparison between ABC and unlabeled antibody PAP procedures. J Histochem Cytochem 29:577.

    Article  CAS  PubMed  Google Scholar 

  • Inouye S, Hondo R (1990): Microplate hybridization of amplified viral DNA segment. J Clin Microbiol 28(6): 1469–1472.

    CAS  PubMed  Google Scholar 

  • Jablonski E, Moomaw E, Tullis RH, Ruth JL (1986): Preparation of oligodeoxynucleotide-alkaline phosphatase conjugates and their use as hybridization probes. Nucl Acids Res 14:6115–6128.

    Article  CAS  PubMed  Google Scholar 

  • Kafatos FC, Jones CW, Efstradiadis A (1979): Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucl Acids Res 7:1541.

    Article  CAS  PubMed  Google Scholar 

  • Katz ED, Haff LA, Eksteen R (1990): Rapid separation, quantitation and purification of products of polymerase chain reaction by liquid chromatography. J Chromatogr 512:433–444.

    Article  CAS  PubMed  Google Scholar 

  • Keller GH, Huang DP, Manak MM (1989): A sensitive nonisotopic hybridization assay for HIV-1 DNA. Anal Biochem 177(1):27–32.

    Article  CAS  PubMed  Google Scholar 

  • Keller GH, Huang DP, Shih JW, Manak MM (1990): Detection of hepatitis B virus DNA in serum by polymerase chain reaction amplification and microtiter sandwich hybridization. J Clin Microbiol 28(6): 1411–1416.

    CAS  PubMed  Google Scholar 

  • Keller GH, Huang DP, Manak MM (1991): Detection of human immunodeficiency virus type 1 DNA by polymerase chain reaction amplification and capture hybridization in microtiter wells. J Clin Microbiol 29(3):638–641.

    CAS  PubMed  Google Scholar 

  • Kelly DJ, Marana DP, Stover CK, Oaks EV, Carl M (1990): Detection of Rickettsia tsutsugamushi by gene amplification using polymerase chain reaction techniques. Ann NY Acad Sci 590:564–571.

    Article  CAS  PubMed  Google Scholar 

  • Kemp DJ, Smith DB, Foote SJ, Samaras N, Peterson MG (1989): Colorimetric detection of specific DNA segments amplified by polymerase chain reactions. Proc Natl Acad Sci USA 86(7): 2423–2427.

    Article  CAS  PubMed  Google Scholar 

  • Kemp DJ, Churchill MJ, Smith DB, Biggs BA, Foote S J, Peterson MG, Samaras N, Deacon NJ, Doherty R (1990): Simplified colorimetric analysis of polymerase chain reactions: Detection of HIV sequences in AIDS patients. Gene 94(2): 223–228.

    Article  CAS  PubMed  Google Scholar 

  • Langer PR, Waldrop AA, Ward DC (1981): Enzymatic synthesis of biotin-labeled polynucleotides: Novel nucleic acid affinity probes. Proc Natl Acad Sci USA 78:6633–6637.

    Article  CAS  PubMed  Google Scholar 

  • Langer-Safer PR, Levine M, Ward DC (1982): Immunological method for mapping genes on Drosophila chromosomes. Proc Natl Acad Sci USA 79:4381–4385.

    Article  CAS  PubMed  Google Scholar 

  • Lanzillo JJ (1990): Preparation of digoxigenin-labeled probes by the polymerase chain reaction. Biotechniques 8(6):620

    CAS  PubMed  Google Scholar 

  • Lanzillo JJ (1990): Preparation of digoxigenin-labeled probes by the polymerase chain reaction. Biotechniques 8(6): 622.

    Google Scholar 

  • Leary JJ, Brigati DJ, Ward DC (1983): 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 USA 80:4045–4049.

    Article  CAS  PubMed  Google Scholar 

  • Letsinger RL, Schott ME (1981): Selectivity in binding a phenanthridinium-dinucleotide derivative to homopolynucleotides. J Amer Chem Soc 103:7394.

    Article  CAS  Google Scholar 

  • Lundeberg J, Wahlberg J, Holmberg M, Pettersson U, Uhlen M (1990): Rapid colorimetric detection of in vitro amplified DNA sequences. DNA Cell Biol 9(4):287–292.

    Article  CAS  PubMed  Google Scholar 

  • Lundeberg J, Wahlberg J, Uhlen M (1991): Rapid colorimetric quantification of PCR-amplified DNA. Biotechniques 10(1):68–75.

    CAS  PubMed  Google Scholar 

  • Mantero G, Zonaro A, Albertini A, Bertolo P, Primi D (1991): DNA enzyme immunoassay: General method for detecting products of polymerase chain reaction. Clin Chem 37(3):422–429.

    CAS  PubMed  Google Scholar 

  • McBride LJ, Koepf SM, Gibbs RA, Salser W, Mayrand PE, Hunkapiller MW, Kronick MN (1989): Automated DNA sequencing methods involving polymerase chain reaction. Clin Chem 35(11): 2196–2201.

    CAS  PubMed  Google Scholar 

  • Morrison LE, Halder TC, Stols LM (1989): Solution-phase detection of polynucleotides using interacting fluorescent labels and competitive hybridization. Anal Biochem 183(2):231–244.

    Article  CAS  PubMed  Google Scholar 

  • Ou CY, McDonough SH, Cabanas D, Ryder TB, Harper M, Moore J, Schochetman G (1990): Rapid and quantitative detection of enzymatically amplified HIV-1 DNA using chemiluminescent oligonucleotide probes. AIDS Res Hum Retrovirus 6(11): 1323–1329.

    CAS  Google Scholar 

  • Pritchard CG, Stefano JE (1990): Amplified detection of viral nucleic acid at subattomole levels using Q beta replicase. Ann Biol Clin (Paris) 48(7):492–497.

    CAS  Google Scholar 

  • Renz M, Kurz C (1984): A colorimetric method for DNA hybridization. Nucl Acids Res 12:3435–3444.

    Article  CAS  PubMed  Google Scholar 

  • Rimstad E, Hornes E, Olsvik O, Hyllseth B (1990): Identification of a double-stranded RNA virus by using polymerase chain reaction and magnetic separation of the synthesized DNA segments. J Clin Microbiol 28(10):2275–2278.

    CAS  PubMed  Google Scholar 

  • Ruth JL (1984): Chemical synthesis of non-radioactively-labeled DNA hybridization probes. DNA 3:123.

    Google Scholar 

  • Ruth JL, Bryan RN (1984): Chemical synthesis of modified oligonucleotides and their utility as non-radioactive hybridization probes. Fed Proc 43:2048 (abst. 3666).

    Google Scholar 

  • Said JW, Sassoon AF, Shintaku IP, Corcoran P, Nichols SW (1990): Polymerase chain reaction for bcl-2 in diagnostic lymph node biopsies. Mod Pathol 3(6):659–663.

    CAS  PubMed  Google Scholar 

  • Saiki RK, Chang CA, Levenson CH, Warren TC, Boehm CD, Kazazian HH Jr, Erlich HA (1988): Diagnosis of sickle cell anemia and beta-thalassemia with enzymatically amplified DNA and nonradioactive allele-specific oligonucleotide probes. N Engl J Med 319(9):537–541.

    Article  CAS  PubMed  Google Scholar 

  • Saiki RK, Walsh PS, Levenson CH, Erlich HA (1989): Genetic analysis of amplified DNA with immobilized sequence-specific oligonucleotide probes. Proc Natl Acad Sci USA 86(16):6230–6234.

    Article  CAS  PubMed  Google Scholar 

  • Scharf SJ, Griffith RL, Erlich HA (1991): Rapid typing of DNA sequence polymorphism at the HLA-DRB1 locus using the polymerase chain reaction and nonradioactive oligonucleotide probes. Hum Immunol 30(3): 190–201.

    Article  CAS  PubMed  Google Scholar 

  • Seibl R, Holtke HJ, Ruger R, Meindl A, Zachau HG, Rasshofer R, Roggendorf M, Wolf H, Arnold N, Wienberg J et al (1990): Non-radioactive labeling and detection of nucleic acids. III. Applications of the digoxigenin system. Biol Chem Hoppe Seyler 371(10):939–951.

    Article  CAS  PubMed  Google Scholar 

  • Smith LM, Fung S, Hunkapiller MW, Hunkapiller TJ, Hood LE (1985): The synthesis of oligonucleotides containing an aliphatic amino group at the 5′ terminus: Synthesis of fluorescent DNA primers for use in DNA sequence analysis. Nucl Acids Res 13:2399–2412.

    Article  CAS  PubMed  Google Scholar 

  • Soderlund H (1990): DNA hybridization: Comparison of liquid and solid phase formats. Ann Biol Clin (Paris) 48(7):489–491.

    CAS  Google Scholar 

  • Sternberger LA, Hardy PH, Cuculis JJ, Meyer HG (1970): The unlabeled antibody-enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333.

    Article  CAS  PubMed  Google Scholar 

  • Sullivan KM, Hopgood R, Lang B, Gill P (1991): Automated amplification and sequencing of human mitochondrial DNA. Electrophoresis 12(1): 17–21.

    Article  CAS  PubMed  Google Scholar 

  • Szabo P, Ward DC (1983): What’s new with hybridization in situ, Tech Biol Sci, 425–427.

    Google Scholar 

  • Syvanen AC, Aalto-Setala K, Kontula K, Soderlund H (1989): Direct sequencing of affinity-captured amplified human DNA application to the detection of apolipoprotein E polymorphism. FEBS Lett 258(1):71–74.

    Article  CAS  PubMed  Google Scholar 

  • Syvanen AC, Aalto-Setala K, Harju L, Kontula K, Soderlund H (1990): A primer-guided nucleotide incorporation assay in the genotyping of apolipoprotein E. Genomics 8(4):684–692.

    Article  CAS  PubMed  Google Scholar 

  • Teo CG, Griffin BE (1990): Visualization of single copies of the Epstein-Barr virus genome by in situ hybridization. Anal Biochem 186(1):78–85.

    Article  CAS  PubMed  Google Scholar 

  • Wahlberg J, Lundeberg J, Hultman T, Holmberg M, Uhlen M (1990a): Rapid detection and sequencing of specific in vitro amplified DNA sequences using solid phase methods. Mol Cell Probes 4(4):285–297.

    Article  CAS  PubMed  Google Scholar 

  • Wahlberg J, Lundeberg J, Hultman T, Uhlen M (1990b): General colorimetric method for DNA diagnostics allowing direct solid-phase genomic sequencing of the positive samples. Proc Natl Acad Sci USA 87(17):6569–6573.

    Article  CAS  PubMed  Google Scholar 

  • Ward DC (1982): Modified nucleosides and methods of preparing and using same. European Patent Office Application Number 82311804.9.

    Google Scholar 

  • Zhang Y, Coyne MY, Will SG, Levenson CH, Kawasaki ES (1991): Single-base mutational analysis of cancer and genetic diseases using membrane bound modified oligonucleotides. Nucl Acids Res 19(14):3929–3933.

    Article  CAS  PubMed  Google Scholar 

Download references

Authors
  1. Richard H. Tullis
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. La Jolla, CA, 92037, USA

    Kary B. Mullis

  2. The Immune Response Corporation, 5935 Darwin Court, Carlsbad, CA, 92008, USA

    François Ferré

  3. Institute for Molecular Genetics, Baylor College of Medicine, One Baylor Plaza, Houston, TX, 77030, USA

    Richard A. Gibbs

Rights and permissions

Reprints and permissions

Copyright information

© 1994 Springer Science+Business Media New York

About this chapter

Cite this chapter

Tullis, R.H. (1994). Ultrasensitive Nonradioactive Detection of PCR Reactions: An Overview. In: Mullis, K.B., Ferré, F., Gibbs, R.A. (eds) The Polymerase Chain Reaction. Birkhäuser, Boston, MA. https://doi.org/10.1007/978-1-4612-0257-8_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-4612-0257-8_10

  • Publisher Name: Birkhäuser, Boston, MA

  • Print ISBN: 978-0-8176-3750-7

  • Online ISBN: 978-1-4612-0257-8

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation