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Development and evaluation of a one-step real-time RT-PCR assay for universal detection of influenza A viruses from avian and mammal species
- Source :
- Archives of Virology
- Publication Year :
- 2010
-
Abstract
- The objective of our study was to develop and evaluate a TaqMan real-time RT-PCR (RRT-PCR) assay for universal detection of influenza A (IA) viruses. The primers and LNA-modified octanucleotide probe were selected to correspond to extremely conserved regions of the membrane protein (MP) segment identified by a comprehensive bioinformatics analysis including 10,405 IA viruses MP sequences, i.e., all of the sequences of the Influenza Virus Sequence database collected as of August 20, 2009. The RRT-PCR has a detection limit of approximately five copies of target RNA/reaction and excellent reaction parameters tested in four IA viruses reference laboratories. The inclusivity of the assay was estimated at both the bioinformatic and the experimental level. Our results predicted that this RRT-PCR assay was able to detect 99.5% of known human IA virus strains, 99.84% of pandemic influenza A (H1N1) strains, 99.75% of avian strains, 98.89% of swine strains, 98.15% of equine strains, and 100% of influenza A viruses of other origin. Electronic supplementary material The online version of this article (doi:10.1007/s00705-010-0636-x) contains supplementary material, which is available to authorized users.
- Subjects :
- medicine.medical_specialty
Swine
Pandemic Influenza
Biology
medicine.disease_cause
Total Nucleic Acid Extraction
Virus
Microbiology
Birds
Medical microbiology
Virology
medicine
TaqMan
Influenza A virus
Animals
Humans
Horses
Sequence database
Reverse Transcriptase Polymerase Chain Reaction
General Medicine
Influenza A virus subtype H5N1
Influenza
Reverse transcription polymerase chain reaction
Real-time polymerase chain reaction
Classical Swine Fever Virus
Original Article
Avian Influenza
Subjects
Details
- ISSN :
- 14328798
- Volume :
- 155
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Archives of virology
- Accession number :
- edsair.doi.dedup.....ecf9ec258390e3103146755b8845b00c