Improved detection of rhinoviruses by nucleic acid sequence-based amplification after nucleotide sequence determination of the 5' noncoding regions of additional rhinovirus strains.
The isothermal nucleic acid sequence-based amplification (NASBA) system was applied for the detection of rhinoviruses using primers targeted at the 5' noncoding region (5' NCR) of the viral genome. The nucleotide sequence of the 5' NCRs of 34 rhinovirus isolates was determined to map the most conserved regions and design more appropriate primers and probes. The assay amplified RNA extracted from 30 rhinovirus reference strains and 88 rhinovirus isolates, it did not amplify RNA from 49 enterovirus isolates and other respiratory viruses. The assay allows one to discriminate between group A and B rhinoviruses. Sensitivities for the detection of group B and group A rhinoviruses was 20 and 200 50% tissue culture infective doses, respectively.
Department of Microbiology, University of Antwerp UIA, Antwerp, Belgium. katherine.loens@ua.ac.be
K
K
Loens
M
M
Ieven
D
D
Ursi
C
C
De Laat
P
P
Sillekens
P
P
Oudshoorn
H
H
Goossens
D020121:5' Untranslated Regions; D001431:Bacteriological Techniques / Q000379:methods* / Q000706:statistics & numerical data; D001483:Base Sequence; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D016679:Genome, Viral; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods* / Q000706:statistics & numerical data; D010850:Picornaviridae Infections / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D012229:Rhinovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid
D020121:5' Untranslated Regions; D017931:DNA Primers; D015342:DNA Probes; D012367:RNA, Viral