Enzyme-linked oligosorbent assay for detection of polymerase chain reaction-amplified human immunodeficiency virus type 1
Autor: | P Cros, E Chapuis, François Mallet, Bernard Mandrand, P Allibert, C Hebrard, D Brand, Francis Barin, J M Besnier |
---|---|
Rok vydání: | 1993 |
Předmět: |
Microbiology (medical)
Genes Viral viruses Microgram Molecular Sequence Data Human immunodeficiency virus (HIV) Molecular Probe Techniques Oligonucleotide Primer HIV Infections Biology medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity law.invention chemistry.chemical_compound law medicine Humans Nucleotide Polymerase chain reaction chemistry.chemical_classification Base Sequence Oligonucleotide Reproducibility of Results virus diseases Virology Molecular biology Enzyme chemistry DNA Viral HIV-1 Oligonucleotide Probes DNA Research Article |
Zdroj: | Journal of Clinical Microbiology. 31:1444-1449 |
ISSN: | 1098-660X 0095-1137 |
Popis: | An enzyme-linked oligosorbent assay (ELOSA) was developed for the detection on microtiter plates of polymerase chain reaction (PCR)-amplified human immunodeficiency virus type 1 (HIV-1) DNA. The denatured PCR product was hybridized with a passively adsorbed oligonucleotide capture probe and a horseradish peroxidase-labeled oligonucleotide detection probe. The sensitivity and specificity of the PCR-ELOSA technique depended to some extent on the nucleotide sequences of the oligonucleotide primer and probe quartet used in the amplification and detection. We evaluated five oligonucleotide quartets located in the gag, pol, vpr, env, and nef regions of HIV-1. DNAs from 39 HIV-1-seropositive individuals and 27 healthy HIV-1-seronegative controls were amplified by the PCR procedure, and the products were detected by ELOSA. Ten copies of HIV-1 DNA against a background of 1 microgram of human DNA were specifically detected by PCR-ELOSA. Specificities and sensitivities were, respectively, 100 and 95% for the gag system, 100 and 97% for the pol system, 100 and 85% for the vpr system, 96 and 95% for the env system, and 100 and 95% for the nef system. The simplicity of ELOSA makes it suitable for automation and applicable to genetic testing and detection of viral and bacterial DNAs or RNAs in most routine laboratories. |
Databáze: | OpenAIRE |
Externí odkaz: |