Nucleic acid amplification in vitro: Detection of sequences with low copy numbers and application to diagnosis of human immunodeficiency virus type 1 infection

Guatelli, J. C., Gingeras, T. R., Richman, D. D. (1989) Nucleic acid amplification in vitro: Detection of sequences with low copy numbers and application to diagnosis of human immunodeficiency virus type 1 infection. Clinical Microbiology Reviews, 2 (2). pp. 217-226. ISSN 08938512 (ISSN)

DOI: 10.1128/CMR.2.2.217


The enzymatic amplification of specific nucleic acid sequences in vitro has revolutionized the use of nucleic acid hybridization assays for viral detection. With this method, the copy number of a pathogen-specific sequence is increased several orders of magnitude before detection is attempted. The sensitivity and specificity of detection are thus markedly improved. Mullis and Faloona devised the first method of sequence amplification in vitro, the polymerase chain reaction (K.B. Mullis and F.A. Faloona, Methods Enzymol. 155:350-355, 1987). By this method, synthetic oligonucleotide primers direct repeated, target-specific, deoxyribonucleic acid-synthetic reactions, resulting in an exponential increase in the amount of the specific target sequence. The application of sequence amplification to viral detection was initially performed with human immunodeficiency virus type 1 and human T-cell lymphoma virus type I. In principle, however, this approach can be applied to the detection of any deoxyribonucleic or ribonucleic acid virus; the only requirement is that sufficient nucleotide sequence data exist to allow the synthesis of target-specific oligonucleotide primers. The use of target amplification in vitro will permit a variety of studies of viral pathogenesis which have not been feasible because of the low copy number of the viral nucleic acids in infected material. This approach is particularly applicable to the study of human retroviral infections, which are chronic and persistent and are characterized by low titers of virus in tissues. In addition, target amplification in vitro will facilitate the development of new methods of sequence detection, which will be useful for rapid viral diagnosis in the clinical laboratory.

Item Type: Paper
Additional Information:
Uncontrolled Keywords: rna cloning gene amplification human human immunodeficiency virus 1 in vitro study lymphoma virus methodology nucleic acid hybridization nucleotide sequence polymerase chain reaction review virus detection virus infection virus pathogenesis virus transcription Acquired Immunodeficiency Syndrome Base Sequence DNA Probes DNA, Viral DNA Directed DNA Polymerase HIV-1 Human T lymphotropic virus 1
Subjects: bioinformatics > genomics and proteomics > annotation > sequence annotation
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene amplification
organism description > virus
CSHL Authors:
Communities: CSHL labs > Gingeras lab
Depositing User: CSHL Librarian
Date: 1989
Date Deposited: 14 Mar 2012 14:20
Last Modified: 31 Mar 2014 13:41
PMCID: PMC358112
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