Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick
Abstract Background Lawsonia intracellularis (L. intracellularis) is the etiologic agent of porcine proliferative enteropathy (PPE), which is reported in many swine breeding countries all over the world, and has caused enormous economic losses in intensive pig production systems. Therefore, the aim...
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doaj-fe89f34f3ca444da99a8c46053f226df2020-11-25T02:56:30ZengBMCBMC Veterinary Research1746-61482019-03-0115111010.1186/s12917-019-1841-9Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstickYanyang Wu0Kaiyue Tian1Yuhan Zhang2Huifang Guo3Ning Li4Zeng Wang5Jun Zhao6College of Animal Science and Veterinary Medicine, Henan Agricultural UniversityCollege of Animal Science and Veterinary Medicine, Henan Agricultural UniversityCollege of Animal Science and Veterinary Medicine, Henan Agricultural UniversityCollege of Animal Science and Veterinary Medicine, Henan Agricultural UniversityCollege of Animal Science and Veterinary Medicine, Henan Agricultural UniversityCollege of Animal Science and Veterinary Medicine, Henan Agricultural UniversityCollege of Animal Science and Veterinary Medicine, Henan Agricultural UniversityAbstract Background Lawsonia intracellularis (L. intracellularis) is the etiologic agent of porcine proliferative enteropathy (PPE), which is reported in many swine breeding countries all over the world, and has caused enormous economic losses in intensive pig production systems. Therefore, the aim of this study was to develop a simple and rapid method for on-site detection of Lawsonia intracellularis (L. intracellularis). As the isothermal recombinase polymerase amplification (RPA) can be performed at a constant temperature and its product is directly observed on a lateral-flow dipstick (LFD) with naked eyes without electrophoresis, the RPA-LFD assay should be useful for field diagnosis of L. intracellularis as well as its detection from clinical samples. Results The established RPA-LFD assay could be finished in 30 min at a wide temperature range of 25 to 40 °C, and the amplicons could be visualized by naked eyes. The developed RPA-LFD assay was specific to dnaA gene of L. intracellularis, and did not detect nucleic acids extracted from other common gastrointestinal pathogens. The minimum detection of this RPA-LFD method was 400 L. intracellularis per reaction, which was as sensitive as conventional PCR. Further, the RPA-LFD assay was performed with 150 clinical fecal samples and the detection results were compared with conventional PCR. Results showed that the coincidence rate of RPA-LFD and conventional PCR was 100%. Conclusions The combined RPA with LFD assay provides a simple, rapid, specific and sensitive alternative for field diagnosis of L. intracellularis infection.http://link.springer.com/article/10.1186/s12917-019-1841-9Lawsonia intracellularisRecombinase polymerase amplificationLateral flow dipstickOn-site detection |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yanyang Wu Kaiyue Tian Yuhan Zhang Huifang Guo Ning Li Zeng Wang Jun Zhao |
spellingShingle |
Yanyang Wu Kaiyue Tian Yuhan Zhang Huifang Guo Ning Li Zeng Wang Jun Zhao Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick BMC Veterinary Research Lawsonia intracellularis Recombinase polymerase amplification Lateral flow dipstick On-site detection |
author_facet |
Yanyang Wu Kaiyue Tian Yuhan Zhang Huifang Guo Ning Li Zeng Wang Jun Zhao |
author_sort |
Yanyang Wu |
title |
Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick |
title_short |
Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick |
title_full |
Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick |
title_fullStr |
Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick |
title_full_unstemmed |
Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick |
title_sort |
rapid and visual detection of lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick |
publisher |
BMC |
series |
BMC Veterinary Research |
issn |
1746-6148 |
publishDate |
2019-03-01 |
description |
Abstract Background Lawsonia intracellularis (L. intracellularis) is the etiologic agent of porcine proliferative enteropathy (PPE), which is reported in many swine breeding countries all over the world, and has caused enormous economic losses in intensive pig production systems. Therefore, the aim of this study was to develop a simple and rapid method for on-site detection of Lawsonia intracellularis (L. intracellularis). As the isothermal recombinase polymerase amplification (RPA) can be performed at a constant temperature and its product is directly observed on a lateral-flow dipstick (LFD) with naked eyes without electrophoresis, the RPA-LFD assay should be useful for field diagnosis of L. intracellularis as well as its detection from clinical samples. Results The established RPA-LFD assay could be finished in 30 min at a wide temperature range of 25 to 40 °C, and the amplicons could be visualized by naked eyes. The developed RPA-LFD assay was specific to dnaA gene of L. intracellularis, and did not detect nucleic acids extracted from other common gastrointestinal pathogens. The minimum detection of this RPA-LFD method was 400 L. intracellularis per reaction, which was as sensitive as conventional PCR. Further, the RPA-LFD assay was performed with 150 clinical fecal samples and the detection results were compared with conventional PCR. Results showed that the coincidence rate of RPA-LFD and conventional PCR was 100%. Conclusions The combined RPA with LFD assay provides a simple, rapid, specific and sensitive alternative for field diagnosis of L. intracellularis infection. |
topic |
Lawsonia intracellularis Recombinase polymerase amplification Lateral flow dipstick On-site detection |
url |
http://link.springer.com/article/10.1186/s12917-019-1841-9 |
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