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Development and evaluation of rapid detection of classical swine fever virus by reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Paper ID Volume ID Publish Year Pages File Format Full-Text
24382 43511 2010 4 PDF Available
Title
Development and evaluation of rapid detection of classical swine fever virus by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
Abstract

A one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed for the detection of classical swine fever virus (CSFV) and a set of primers designed based on the E2 gene reference sequences of the CSFV. The assay was optimized to amplify CSFV RNA by incubation at 63 °C for 50 min. The RT-LAMP amplification products had a ladder-like appearance when electrophoresed on an agarose gel. The RT-LAMP assay showed higher sensitivity than the conventional RT-PCR, and no cross-reactivity appeared with other related porcine viruses including porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine pseudorabies virus (PRV), and porcine reproductive and respiratory syndrome virus (PRRSV). The positive predictive value (PPV) of CSFV RT-LAMP for 227 field tissue samples was 94.7%, the negative predictive value (NPV) was 30.8% and showed better for the conventional RT-PCR method.Thus, the RT-LAMP assay is extremely rapid, sensitive, and specific and has potential usefulness for rapid laboratory diagnosis and pen-side detection for CSFV detection in pigs.

Keywords
Classical swine fever virus; Loop-mediated isothermal amplification (LAMP); RT-PCR; E2 gene
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Development and evaluation of rapid detection of classical swine fever virus by reverse transcription loop-mediated isothermal amplification (RT-LAMP)
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 146, Issue 4, 15 April 2010, Pages 147–150
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us