Detection of human immunodeficiency virus type 1 in AIDS patients using amplification-mediated hybridization analyses: Reproducibility and quantitative limitations

Davis, G. R., Blumeyer, K., DiMichele, L. J., Whitfield, K. M., Chappelle, H., Riggs, N., Ghosh, S. S., Kao, P. M., Fahy, E., Kwoh, D. Y., Guatelli, J. C., Spector, S. A., Richman, D. D., Gingeras, T. R. (1990) Detection of human immunodeficiency virus type 1 in AIDS patients using amplification-mediated hybridization analyses: Reproducibility and quantitative limitations. Journal of Infectious Diseases, 162 (1). pp. 13-20. ISSN 00221899 (ISSN)

URL: http://www.ncbi.nlm.nih.gov/pubmed/2113072
DOI: 10.1093/infdis/162.1.13

Abstract

Eighty-six peripheral blood mononuclear cell (PBMC) samples from 30 patients with AIDS were analyzed using a transcription-based amplification system (TAS) and the polymerase chain reaction (PCR). Human immunodeficiency virus tpe 1 (HIV-1) sequences were detected by amplification-mediated hybridization in 98% of the samples, 52% of which were positive for p24 antigen by ELISA. Neither TAS (93%) nor PCR (95%) detected HIV-1 sequences in all 86 samples. The hybridization-detection methods (slot blot, bead-based sandwich, and solution) used to detect the HIV-1-specific TAS products had a clear influence on the efficiency of detecting and quantitating the levels of HIV-1 present in these samples. The reproducibility of amplification of constant amounts of HIV-1 RNA and β-globin DNA by TAS and PCR was studied over 3 months. The results indicated that variations of 10- and 5-fold in the HIV-1 sequence levels could be detected between samples by TAS and PCR, respectively. Within the range of sensitivities for each assay used, the administration of zidovudine did not appear to reduce the amount of HIV-1 nucleic acid sequences as observed in PBMC obtained serially from six AIDS patients.

Item Type: Paper
Uncontrolled Keywords: virus antigen acquired immune deficiency syndrome antibody detection article enzyme linked immunosorbent assay gene amplification human human immunodeficiency virus 1 hybridization mononuclear cell nucleotide sequence polymerase chain reaction priority journal virus detection Acquired Immunodeficiency Syndrome Base Sequence Gene Products, gag HIV Antigens HIV Core Protein p24 HIV-1 Molecular Sequence Data Nucleic Acid Hybridization Predictive Value of Tests Reproducibility of Results Transcription Genetic Viral Core Proteins
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > transcription
diseases & disorders > viral diseases > HIV
Investigative techniques and equipment > cloning > PCR
Investigative techniques and equipment > assays > cloning > PCR

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene amplification
CSHL Authors:
Communities: CSHL labs > Gingeras lab
Depositing User: CSHL Librarian
Date: 1990
Date Deposited: 14 Mar 2012 14:30
Last Modified: 28 Mar 2014 20:54
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25245

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving