+ 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

Development and evaluation of NucliSens basic kit NASBA for diagnosis of parainfluenza virus infection with 'end-point' and 'real-time' detection



Development and evaluation of NucliSens basic kit NASBA for diagnosis of parainfluenza virus infection with 'end-point' and 'real-time' detection



Journal of Virological Methods 108(2): 145-155



New methods for the detection of human parainfluenza viruses (HPIVs) were developed. These were based on nucleic acid sequence-based amplification (NASBA) and utilised the NucliSens Basic Kit. Primers and probes were selected from the haemagglutinin neuraminidase (HN) gene of HPIV1, HPIV2 and HPIV3, and from the phosphoprotein (P) of HPIV4a and -4b. Synthetic RNA, titrated control virus stocks and respiratory specimens (n=44) were utilised to evaluate performance of the assays. Detection of NASBA products was by probe hybridisation and electrochemiluminescence (ECL) ('end-point' detection) or using molecular beacons ('real-time' detection). The assays using ECL detection proved to be both sensitive and specific. Typically, less than or equal to 100 RNA copies or one TCID(50) input was detectable with no cross-reaction between the specific HPIV assays and other respiratory viruses. Results for clinical samples were concordant with those obtained by 'conventional' procedures by classical viral diagnostic methods. 'Real-time' detection utilised probes specific for either HPIV1 or HPIV3 with similar performance characteristics to the assays with 'end-point' detection. The feasibility of multiplexing targets together was confirmed using a combined HPIV1 and HPIV3 assay with good results for ECL and molecular beacon detection on control material and clinical samples.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 048758377

Download citation: RISBibTeXText

PMID: 12609681

DOI: 10.1016/s0166-0934(02)00268-9


Related references

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, 2001

Evaluation of a new NASBA assay for the qualitative detection of hepatitis C virus based on the NucliSens Basic Kit reagents. Journal of Clinical Virology 29(2): 84-91, 2004

Development and evaluation of the Nuclisens Basic Kit NASBA for the detection of RNA from yeast frequently resistant to antifungal drugs. Abstracts of the Interscience Conference on Antimicrobial Agents & Chemotherapy 41: 385, 2001

Development and evaluation of the nuclisens basic kit NASBA for the detection of RNA from Candida species frequently resistant to antifungal drugs. Diagnostic Microbiology & Infectious Disease 45(3): 217-220, 2003

Development and evaluation of nucleic acid sequence based amplification (NASBA) for diagnosis of enterovirus infections using the NucliSens Basic Kit. Journal of Clinical Virology 24(1-2): 117-130, 2002

Evaluation of the NucliSens miniMAG RNA extraction and real-time NASBA applications for the detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae in throat swabs. Journal of Microbiological Methods 72(2): 217-219, 2008

Clinical evaluation of NucliSENS magnetic extraction and NucliSENS analytical specific reagents for the real-time detection of respiratory syncytial virus (RSV) in paediatric respiratory specimens. Journal of Clinical Pathology 62(11): 998, 2009

Evaluation of the Nuclisens basic kit NASBA for the detection of RNA from yeasts frequently resistant to antifungal drugs. Blood 98(11 Part 2): 357b, 2001

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

Development of a real-time RT-PCR assay for detection and quantitation of parainfluenza virus 3. Journal of Virological Methods 130(1-2): 145-148, 2005

Real-time NASBA for detection of Strawberry vein banding virus. European Journal of Plant Pathology 110(2): 213-221, 2004

Comparison of the real-time nucleic acid sequence-based amplification (RTi-NASBA) with conventional NASBA, and galactomannan assay for the diagnosis of invasive aspergillosis. Journal of Korean Medical Science 22(4): 672-676, 2007

Evaluation of the NucliSens Basic Kit assay for detection of Norwalk virus RNA in stool specimens. Journal of Virological Methods 108(1): 123-131, 2003