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Titolo:
Rapid multiserotype detection of human rhinoviruses on optically coated silicon surfaces
Autore:
Ostroff, R; Ettinger, A; La, H; Rihanek, M; Zalman, L; Meador, J; Patick, AK; Worland, S; Polisky, B;
Indirizzi:
Thermo BioStar Inc, Boulder, CO 80301 USA Thermo BioStar Inc Boulder CO USA 80301 ioStar Inc, Boulder, CO 80301 USA Agouron Pharmaceut Inc, San Diego, CA 92121 USA Agouron Pharmaceut Inc San Diego CA USA 92121 nc, San Diego, CA 92121 USA
Titolo Testata:
JOURNAL OF CLINICAL VIROLOGY
fascicolo: 2, volume: 21, anno: 2001,
pagine: 105 - 117
SICI:
1386-6532(200105)21:2<105:RMDOHR>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
UPPER RESPIRATORY-TRACT; 3C PROTEASE; 5'-NONCODING REGION; ANTIVIRAL ACTIVITY; RNA-BINDING; INFECTIONS; SEROTYPES; IMMUNOASSAY; FAMILIES; VIRUS;
Keywords:
rhinovirus detection; optical immunoassay; OIA; thin film biosensor; 3C protease;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Citazioni:
52
Recensione:
Indirizzi per estratti:
Indirizzo: Ostroff, R Thermo BioStar Inc, 6655 Lookout Rd, Boulder, CO 80301 USA Thermo BioStar Inc 6655 Lookout Rd Boulder CO USA 80301 301 USA
Citazione:
R. Ostroff et al., "Rapid multiserotype detection of human rhinoviruses on optically coated silicon surfaces", J CLIN VIRO, 21(2), 2001, pp. 105-117

Abstract

Background: More than 100 immunologically distinct serotypes of human rhinoviruses (HRV) have been discovered, making detection of surface exposed capsid antigens impractical. However, the non-structural protein 3C protease (3Cpro) is essential for viral replication and is relatively highly conserved among serotypes. making it a potential tai get for diagnostic testing. The thin him biosensor is an assay platform that can be formatted into a sensitive immunoassay for viral proteins in clinical specimens. The technologyutilizes an optically coated silicon surface to convert specific molecularbinding events into visual color changes by altering the reflective properties of light through molecular thin films, Objective: To develop a rapid test for detection of HRV by developing broadly serotype reactive antibodiesto 3Cpro and utilizing them in the thin film biosensor format. Study design: Polyclonal antibodies to 3Cpro were purified and incorporated into the thin film assay. The in vitro sensitivity, specificity and multiserotype cross-reactivity of the 3Cpro assay were tested. Nasal washes from naturally infected individuals were also tested to verify that 3Cpro was detectable ill clinical specimens. Results: The 3Cpro assay is: a 28-min, non-instrumented room temperature test with a visual limit of detection of 12 pM (picomolar) 3Cpro. In terms of viral titer, as few as 1000 TCID50 equivalents of HRV2 were detectable. The assay detected 45/52 (87%) of the HRV serotypes tested but showed no cross-reactivity to common respiratory viruses or bacteria. The thin film assay detected 3Cpro ill HRV-infected cell culture supernatants coincident with first appearance of cytopathic effect. Data are also presented demonstrating 3Cpro detection from clinical samples collected from HRV-infected individuals. The assay detected 3Cpro in expelled nasal secretions from a symptomatic individual on the first day of illness. In addition, 9/11 (82%) concentrated nasal wash specimens from HRV infected childrenwere positive in the 3Cpro lest. Conclusion: We have described a novel, sensitive thin film biosensor for rapid detection of HRV 3Cpro. This test maybe suitable for the point of care setting, where rapid HRV diagnostic testresults could contribute to clinical decisions regarding appropriate antibiotic or antiviral therapy. (C) 2001 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/09/20 alle ore 09:05:00