+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

NucliSens Basic Kit NASBA with real-time detection for accelerated laboratory diagnosis of enterovirus infections

NucliSens Basic Kit NASBA with real-time detection for accelerated laboratory diagnosis of enterovirus infections

Abstracts of the General Meeting of the American Society for Microbiology 101: 264

Background: Enteroviruses are among the most common viral pathogens of humans and are responsible for a wide range of clinical symptoms. While culture is useful in the detection of patients with an enterovirus infection, it is time-consuming and lacks sensitivity. Objective: The aim of this study was to develop a molecular diagnostic assay for the detection of enteroviruses in a variety of clinical specimens using the NASBA amplification technology. Methods: Primers and probes were derived from the 5' non-coding region of the enterovirus genome (pan-enterovirus assay). For nucleic acid extraction and subsequent RNA amplification and detection, reagents from the NucliSens Basic Kit were used. Reference enterovirus strains and clinical samples were analyzed. Results: Initially, the assay utilized degenerate primers and 'end-point' detection of amplification products by electrochemiluminescence. The sensitivity of the pan-enterovirus assay, determined on cultures of a range of enteroviruses, was about 1 TCID50. The specificity was 100%, as determined by examination of enterovirus reference strains and non-enteroviruses including genetically closely related rhinoviruses. Prospectively-analyzed clinical samples (cerebrospinal fluids (CSF), respiratory tract samples, and stool specimens) revealed concordant results for NASBA and an 'in-house' RT-PCR. The assay format was simplified and made more user-friendly by incorporating non-degenerate primers for the amplification and a molecular beacon for 'real time' detection. In addition, an internal control RNA was introduced to monitor isolation, amplification and detection at the individual sample level. None of these changes had a major impact on the sensitivity and specificity of the assay. Conclusions: The pan-enterovirus NASBA, in NucliSens Basic Kit format, is a clinically useful assay applicable to a wide range of clinical specimens. Use of an internal control RNA and 'real time' detection enlarges the utility of the assay in a routine diagnostic setting and enables diagnosis of enterovirus infections within a few hours.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 035413812

Download citation: RISBibTeXText

Related references

Development and evaluation of N-hydroxysuccinimide-activated silica for immobilizing human serum albumin in liquid chromatography columns. Journal of Chromatography. a 1049(1-2): 51-61, 2004

Development and evaluation of nasal formulations of ketorolac. Drug Delivery 7(4): 223-229, 2000

Evaluation of the NT-proBNP as precoce biological marker of the cardiac dysfunction in the septic shock FT Evaluation du NT-proBNP comme marqueur biologique de la dysfonction cardiaque du choc septique. 2008

Development and evaluation of multiple-objective decision-making methods for watershed management planning. Journal of the American Water Resources Association 38(2): 517-529, 2002

Evaluation of the Nuclisens CMV pp67 assay for detection of cytomegalovirus infection after allogeneic stem cell transplantation. Abstracts of the Interscience Conference on Antimicrobial Agents and Chemotherapy 39: 438, 1999

Isolation and Characterization of the Bovine Microsatellite Loci. Biochemical genetics 44(11-12): 518-532, 2006

Development and evaluation of the Lake Macroinvertebrate Integrity Index for New Jersey lakes and reservoirs. Environmental Monitoring and Assessment 77(3): 311-333, 2002

Evaluation of a new Internet-based self-help guide for patients with bulimic symptoms in Sweden. Nordic Journal of Psychiatry 60(6): 463-468, 2006

Development and evaluation of the neutralizing capacity of horse antivenom against the Brazilian spider Loxosceles intermedia. Toxicon 37(9): 1323-1328, 1999

Comparison of the NovaSom QSG, a new sleep apnea home-diagnostic system, and polysomnography. Sleep Medicine 4(3): 213-218, 2003

Clinical evaluation of Nigella sativa seeds for the treatment of hyperlipidemia: a randomized, placebo controlled clinical trial. Medical Archives 66(3): 198-200, 2012

Comparison of the reaction sites associated with the resistance of coleoptiles of barley, oats and wheat to infection by Septoria nodorum. Phytopathologische Zeitschrift = Journal of phytopathology 115(1): 72-82, 1986

Clinical evaluation of NucliSENS magnetic extraction and NucliSENS analyte-specific reagents for real-time detection of human metapneumovirus in pediatric respiratory specimens. Journal of Clinical Microbiology 46(4): 1274-1280, 2008