<abstract xmlns="http://www.w3.org/1999/xhtml"><p>A simple and rapid real-time loop-mediated isothermal amplification (LAMP) assay designed to detect <italic>Lawsonia</italic> (<italic>L</italic>.) <italic>intracellularis</italic>, an important bacteria causing proliferative enteropathy in pigs. A set of four primers targeting the ubiquinone/menaquinone biosynthesis methylase (<italic>ubi</italic>E) gene was designed for the LAMP reaction. Additionally, serial 10-fold dilutions of cultured <italic>L. intracellularis</italic> and spiked feces were also used for the optimization of real-time LAMP. The lower limit of the linear range of the assay in <italic>L. intracellularis</italic> was 1.0 × 10<sup>0</sup><italic>L</italic>. <italic>intracellularis</italic>. Real-time LAMP was 10 and 100 times more sensitive than real-time PCR and conventional PCR detection methods, respectively. Based on testing of 213 porcine fecal samples using real-time LAMP, realtime PCR and PCR, the agreement quotients of real-time LAMP with conventional PCR and with real-time PCR were 0.77 and 0.95, respectively. This study demonstrated that real-time LAMP was a powerful tool for the rapid and sensitive detection of <italic>L</italic>. <italic>intracellularis</italic> in porcine fecal samples.</p></abstract>

A simple and rapid real-time loop-mediated isothermal amplification (LAMP) assay designed to detect Lawsonia (L.) intracellularis, an important bacteria causing proliferative enteropathy in pigs. A set of four primers targeting the ubiquinone/menaquinone biosynthesis methylase (ubiE) gene was designed for the LAMP reaction. Additionally, serial 10-fold dilutions of cultured L. intracellularis and spiked feces were also used for the optimization of real-time LAMP. The lower limit of the linear range of the assay in L. intracellularis was 1.0 × 100L. intracellularis. Real-time LAMP was 10 and 100 times more sensitive than real-time PCR and conventional PCR detection methods, respectively. Based on testing of 213 porcine fecal samples using real-time LAMP, realtime PCR and PCR, the agreement quotients of real-time LAMP with conventional PCR and with real-time PCR were 0.77 and 0.95, respectively. This study demonstrated that real-time LAMP was a powerful tool for the rapid and sensitive detection of L. intracellularis in porcine fecal samples.

Rapid And Sensitive Detection Of <italic>Lawsonia Intracellularis In Pigs</italic> By Real-Time Loop-Mediated Isothermal Amplification

College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University, Jeonju 561-756, Republic of Korea

Department of Animal Biotechnology, Chonbuk National University, Jeonju 561-756, Republic of Korea

College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea

Department of Biology, College of Natural Sciences, Chungbuk National University, Cheongju, Chungbuk 361-763, Republic of Korea

Department of Molecular Medicine and Ewha Medical Research Institute, Ewha Womans University Medical School, Seoul 158-710, Republic of Korea

Acta Veterinaria

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Rapid And Sensitive Detection Of  By Real-Time Loop-Mediated Isothermal Amplification

A simple and rapid real-time loop-mediated isothermal amplification (LAMP) assay designed to detect Lawsonia (L.) intracellularis, an important bacteria...

Rapid And Sensitive Detection Of Lawsonia Intracellularis In Pigs By Real-Time Loop-Mediated Isothermal Amplification

Publié en ligne: 20 mars 2015

Pages: 20 - 29

Reçu: 02 août 2014

Accepté: 22 déc. 2014

DOI: https://doi.org/10.1515/acve-2015-0002

Mots clés
pig, porcine proliferative enteropathy, real-time loopmediated isothermal amplification

© PARK Byung-Yong, SHIM Kwan-Seob, KIM Won-Il, HOSSAIN Md Mukter, KIM Bumseok, KWON Jungkee, PARK Choi-Kyu, CHO Sung-Jin, JO Inho, CHO Ho-Seong

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Rapid And Sensitive Detection Of Lawsonia Intracellularis In Pigs By Real-Time Loop-Mediated Isothermal Amplification

Publié en ligne: 20 mars 2015

Pages: 20 - 29

Reçu: 02 août 2014

Accepté: 22 déc. 2014

DOI: https://doi.org/10.1515/acve-2015-0002

Mots cléspig, porcine proliferative enteropathy, real-time loopmediated isothermal amplification

© PARK Byung-Yong, SHIM Kwan-Seob, KIM Won-Il, HOSSAIN Md Mukter, KIM Bumseok, KWON Jungkee, PARK Choi-Kyu, CHO Sung-Jin, JO Inho, CHO Ho-Seong

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Mots clés
pig, porcine proliferative enteropathy, real-time loopmediated isothermal amplification