A Novel RT-LAMP Assay for Rapid and Simple Detection of Classical Swine Fever Virus
A simple and rapid assay for the detection of Classical swine fever virus (CSFV) was established using reverse transcription loop-mediated isothermal amplification (RT-LAMP). This study describes the amplification of the genomic RNA of CSFV under isothermal conditions (63 ℃) within one hour, using a...
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Published in | Virologica Sinica Vol. 25; no. 1; pp. 59 - 64 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English Chinese |
Published |
Heidelberg
SP Wuhan Institute of Virology, CAS
01.02.2010
China Institute of Yeterinary Drug Control,Beijing,100081,China |
Subjects | |
Online Access | Get full text |
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Summary: | A simple and rapid assay for the detection of Classical swine fever virus (CSFV) was established using reverse transcription loop-mediated isothermal amplification (RT-LAMP). This study describes the amplification of the genomic RNA of CSFV under isothermal conditions (63 ℃) within one hour, using a set of six primers (two outer primers, two inner primers and two loop primers). This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR. This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV. PRRSV. SIV PRV-PCV, thus showed a good specificity. Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition, either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye. Because RT-LAMP is low-cost and produces rapid results, it has the potential to be an excellent tool for CSFV surveillance in the field, especially in developing countries |
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Bibliography: | S852.651 TP309 Reverse transcription loop-mediated isothermal amplification(RT-LAMP) 42-1760/Q Classical swine fever virus (CSFV); Reverse transcription loop-mediated isothermal amplification(RT-LAMP); Rapid detection Rapid detection Classical swine fever virus (CSFV) ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Undefined-1 ObjectType-Feature-3 |
ISSN: | 1674-0769 1995-820X |
DOI: | 10.1007/s12250-010-3043-2 |