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Broadly targeted mu...
Broadly targeted multiprobe QPCR for detection of coronaviruses : Coronavirus is common among mallard ducks (Anas platyrhynchos)
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- Muradrasoli, Shaman (author)
- Uppsala universitet,Klinisk virologi,Blomberg,Uppsala University
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- Mohamed, Nahla (author)
- Uppsala universitet,Klinisk virologi,Blomberg
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- Hornyák, Akos (author)
- SVA
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- Fohlman, Jan (author)
- Växjö Universitet
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- Olsen, Björn (author)
- Uppsala universitet,Infektionssjukdomar,Olsen
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- Belak, Sandor (author)
- Swedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Inst för biomedicin och veterinär folkhälsovetenskap,Department of Biomedical Science and Veterinary Public Health,SVA
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- Blomberg, Jonas (author)
- Uppsala universitet,Klinisk virologi,blomberg
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(creator_code:org_t)
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- Elsevier BV, 2009
- 2009
- English.
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In: Journal of Virological Methods. - : Elsevier BV. - 0166-0934 .- 1879-0984. ; 159:2, s. 277-287
- Related links:
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https://doi.org/10.1...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://res.slu.se/i...
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Abstract
Subject headings
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- Coronaviruses (CoV) can cause trivial or fatal disease in humans and in animals. Detection methods for a wide range of CoVs are needed, to understand viral evolution, host range, transmission and maintenance in reservoirs. A new concept, "Multiprobe QPCR", which uses a balanced mixture of competing discrete non- or moderately degenerated nuclease degradable (TaqMan((R))) probes was employed. It provides a broadly targeted and rational single tube real-time reverse transcription PCR ("NQPCR") for the generic detection and discovery of CoV. Degenerate primers, previously published, and the new probes, were from a conserved stretch of open reading frame 1b, encoding the replicase. This multiprobe design reduced the degree of probe degeneration, which otherwise decreases the sensitivity, and allowed a preliminary classification of the amplified sequence directly from the QPCR trace. The split probe strategy allowed detection of down to 10 viral nucleic acid equivalents of CoV from all known CoV groups. Evaluation was with reference CoV strains, synthetic targets, human respiratory samples and avian fecal samples. Infectious-Bronchitis-Virus (IBV)-related variants were found in seven of 35 sample pools, from 100 wild mallards (Anas platyrhynchos). Ducks may spread and harbour CoVs. NQPCR can detect a wide range of CoVs, as illustrated for humans and ducks.
Subject headings
- LANTBRUKSVETENSKAPER -- Veterinärmedicin (hsv//swe)
- AGRICULTURAL SCIENCES -- Veterinary Science (hsv//eng)
- LANTBRUKSVETENSKAPER -- Husdjursvetenskap (hsv//swe)
- AGRICULTURAL SCIENCES -- Animal and Dairy Sience (hsv//eng)
Keyword
- Coronaviruses
- Real-time PCR
- Nuclease-based probes
- TaqMan
- Avian coronavirus
- Split probe strategy
- MEDICINE
- MEDICIN
- Medical Virology
- Medicinsk virologi
Publication and Content Type
- ref (subject category)
- art (subject category)
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