Development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assay
Abstract Background Porcine epidemic diarrhoea (PED) is an emerging disease in pigs that causes massive economic losses in the swine industry, with high mortality in suckling piglets. Early identification of PED virus (PEDV)-infected herd through surveillance or monitoring strategies is necessary fo...
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doaj-def182ada60e495d93abe48662278bcc2020-11-25T02:12:18ZengBMCBMC Veterinary Research1746-61482018-05-011411810.1186/s12917-018-1498-9Development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assayThi Ngan Mai0Van Diep Nguyen1Wataru Yamazaki2Tamaki Okabayashi3Shuya Mitoma4Kosuke Notsu5Yuta Sakai6Ryoji Yamaguchi7Junzo Norimine8Satoshi Sekiguchi9Animal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAnimal Infectious Disease and Prevention, Department of Veterinary Sciences, Faculty of Agriculture, University of MiyazakiAbstract Background Porcine epidemic diarrhoea (PED) is an emerging disease in pigs that causes massive economic losses in the swine industry, with high mortality in suckling piglets. Early identification of PED virus (PEDV)-infected herd through surveillance or monitoring strategies is necessary for mass control of PED. However, a common working diagnosis system involves identifying PEDV-infected animals individually, which is a costly and time-consuming approach. Given the above information, the thrusts of this study were to develop a real-time fluorescent reverse transcription loop-mediated isothermal amplification (RtF-RT-LAMP) assay and establish a pooled testing system using faecal sample to identify PEDV-infected herd. Results In this study, we developed an accurate, rapid, cost-effective, and simple RtF- RT-LAMP assay for detecting the PEDV genome targeting M gene. The pooled testing system using the RtF-RT-LAMP assay was optimized such that a pool of at least 15 individual faecal samples could be analysed. Conclusions The developed RtF-RT-LAMP assay in our study could support the design and implementation of large-scaled epidemiological surveys as well as active surveillance and monitoring programs for effective control of PED.http://link.springer.com/article/10.1186/s12917-018-1498-9PEDVRtF-RT-LAMPOne-step RT-PCRPooled stool samples |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Thi Ngan Mai Van Diep Nguyen Wataru Yamazaki Tamaki Okabayashi Shuya Mitoma Kosuke Notsu Yuta Sakai Ryoji Yamaguchi Junzo Norimine Satoshi Sekiguchi |
spellingShingle |
Thi Ngan Mai Van Diep Nguyen Wataru Yamazaki Tamaki Okabayashi Shuya Mitoma Kosuke Notsu Yuta Sakai Ryoji Yamaguchi Junzo Norimine Satoshi Sekiguchi Development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assay BMC Veterinary Research PEDV RtF-RT-LAMP One-step RT-PCR Pooled stool samples |
author_facet |
Thi Ngan Mai Van Diep Nguyen Wataru Yamazaki Tamaki Okabayashi Shuya Mitoma Kosuke Notsu Yuta Sakai Ryoji Yamaguchi Junzo Norimine Satoshi Sekiguchi |
author_sort |
Thi Ngan Mai |
title |
Development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assay |
title_short |
Development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assay |
title_full |
Development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assay |
title_fullStr |
Development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assay |
title_full_unstemmed |
Development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assay |
title_sort |
development of pooled testing system for porcine epidemic diarrhoea using real-time fluorescent reverse-transcription loop-mediated isothermal amplification assay |
publisher |
BMC |
series |
BMC Veterinary Research |
issn |
1746-6148 |
publishDate |
2018-05-01 |
description |
Abstract Background Porcine epidemic diarrhoea (PED) is an emerging disease in pigs that causes massive economic losses in the swine industry, with high mortality in suckling piglets. Early identification of PED virus (PEDV)-infected herd through surveillance or monitoring strategies is necessary for mass control of PED. However, a common working diagnosis system involves identifying PEDV-infected animals individually, which is a costly and time-consuming approach. Given the above information, the thrusts of this study were to develop a real-time fluorescent reverse transcription loop-mediated isothermal amplification (RtF-RT-LAMP) assay and establish a pooled testing system using faecal sample to identify PEDV-infected herd. Results In this study, we developed an accurate, rapid, cost-effective, and simple RtF- RT-LAMP assay for detecting the PEDV genome targeting M gene. The pooled testing system using the RtF-RT-LAMP assay was optimized such that a pool of at least 15 individual faecal samples could be analysed. Conclusions The developed RtF-RT-LAMP assay in our study could support the design and implementation of large-scaled epidemiological surveys as well as active surveillance and monitoring programs for effective control of PED. |
topic |
PEDV RtF-RT-LAMP One-step RT-PCR Pooled stool samples |
url |
http://link.springer.com/article/10.1186/s12917-018-1498-9 |
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