Journal article
Enzymatic Gene Amplification: Qualitative and Quantitative Methods for Detecting Proviral DNA Amplified in Vitro
The Journal of infectious diseases, v 158(6), pp 1158-1169
Dec 1988
PMID: 3198934
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
We evaluated various detection methods to identify amplified human retroviral sequences after Thermus aquaticus-directed polymerase chain reaction (PCR). A combination of hybridization formats and direct incorporation assays provided the most information. This multiphasic approach enabled us to detect specific human T cell leukemia virus type I (HTLV-I)-homologous regions in several HTLV-I-seronegative patients with T cell lymphoma, as well as variants of HTLV-I and human immunodeficiency virus type 1 in patients with prototype disease. In all diagnostic assays designed to detect a particular retrovirus, it was necessary to include a hybridization step, because sequences (endogenous or exogenous) homologous to certain primers were present in most human DNA preparations and yielded discrete products, sometimes of the predicted molecular weight, after amplification. These products could be discriminated by hybridization from amplified prototype proviral sequences. The intensity of the signal generated after hybridization was proportional to input target DNA, an observation making it feasible to quantitatively measure the proviral load in a DNA sample.
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Details
- Title
- Enzymatic Gene Amplification: Qualitative and Quantitative Methods for Detecting Proviral DNA Amplified in Vitro
- Creators
- Mark A. Abbott - Department of Medicine, SUNY Health Science Center 13210.Bernard J. Poiesz - Division of Hematology/Oncology, Department of Medicine, SUNY Health Science Center at Syracuse, Syracuse, New York, Syracuse, New York, Emeryville, CaliforniaBruce C. Byrne - Division of Hematology/Oncology, Department of Medicine, SUNY Health Science Center at Syracuse, Syracuse, New York, Syracuse, New York, Emeryville, CaliforniaShirley Kwok - Division of Hematology/Oncology, Department of Medicine, SUNY Health Science Center at Syracuse, Syracuse, New York, Syracuse, New York, Emeryville, CaliforniaJohn J. Sninsky - Division of Hematology/Oncology, Department of Medicine, SUNY Health Science Center at Syracuse, Syracuse, New York, Syracuse, New York, Emeryville, CaliforniaGarth D. Ehrlich - Division of Hematology/Oncology, Department of Medicine, SUNY Health Science Center at Syracuse, Syracuse, New York, Syracuse, New York, Emeryville, California
- Publication Details
- The Journal of infectious diseases, v 158(6), pp 1158-1169
- Publisher
- The University Chicago Press
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:A1988R227100002
- Scopus ID
- 2-s2.0-0024213634
- Other Identifier
- 991019196437804721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Immunology
- Infectious Diseases
- Microbiology