alexa Rapid detection of H5 avian influenza virus by TaqMan-MGB real-time RT-PCR
Infectious Diseases

Infectious Diseases

Journal of Infectious Diseases & Therapy

Author(s): YY Lu, JY Yan, Y Feng, CP Xu, W Shi

Abstract Share this page

AIMS: Real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on a TaqMan-minor groove binder (MGB) probe was developed for the rapid detection of avian influenza virus subtype H5. METHODS AND RESULTS: Conserved regions in the haemagglutinin genes of avian influenza viruses subtype H5 served as targets for the primers and TaqMan-MGB probe design. Concentrations of primers and probe were optimized to improve the sensitivity and specificity of the reactions. A plasmid containing the haemagglutinin gene was constructed and in vitro transcribed for a quantitative assay of copy numbers of the target gene. The results revealed that the optimal concentration of primers and probe was 640 and 480 nmol l(-1), respectively. The threshold of 100 copies of target molecules could be detected. The linear range for detection was determined as 10(2) to 10(8) molecules in reaction. CONCLUSIONS: It took less than 3 h to complete the detection from viral RNA extraction, with good sensitivity and repeatability. SIGNIFICANCE AND IMPACT OF THE STUDY: Real-time RT-PCR assay with MGB probe was an effective means for quick and quantitative laboratory detection and monitoring of H5 avian influenza viruses.

This article was published in Letters in Applied Microbiology and referenced in Journal of Infectious Diseases & Therapy

Relevant Expert PPTs

Relevant Speaker PPTs

Recommended Conferences

Relevant Topics

Peer Reviewed Journals
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version