Real-time loop-mediated isothermal amplification (LAMP) of mgc2 gene of Mycoplasma gallisepticum
| Autor: | Syed Ehtisham-ul-Haque, Madiha Kiran, Usman Waheed, Muhammad Younus |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Mycoplasma gallisepticum 040301 veterinary sciences Veterinary medicine Mycoplasma species Loop-mediated isothermal amplification real-time lamp medicine.disease_cause 0403 veterinary science 03 medical and health sciences SF600-1100 medicine Gene mgc2 gene General Veterinary biology poultry Potential field 04 agricultural and veterinary sciences Mycoplasma biology.organism_classification Virology Agglutination (biology) 030104 developmental biology Gene sequence mycoplasma gallisepticum Research Article |
| Zdroj: | Journal of Veterinary Research, Vol 61, Iss 4, Pp 439-444 (2017) Journal of Veterinary Research |
| ISSN: | 2450-8608 |
| Popis: | Introduction: Mycoplasma gallisepticum is considered the most pathogenic and economically significant avian Mycoplasma spp. for the worldwide poultry industry. The aim of this study was to develop a novel and sensitive real-time loop-mediated isothermal amplification (LAMP) assay based on the amplification of its mgc2 gene sequence for its rapid molecular detection in poultry. Material and Methods: Blood samples from 300 broiler and layer chickens were screened using a rapid serum agglutination (RSA) test. A real-time LAMP reaction was conducted with seropositive swab samples at 60°C for 90 min in an ESEQuant tube scanner using 6-carboxyfluorescein as the reporting dye. Results: The sensitivity of the developed assay was 10 fg/μL of DNA. The assay was found 100% specific, showing no cross-reactivity with other avian Mycoplasma species. The proportion found of the positive samples by the real-time LAMP was 58%. In comparison, the RSA was found to detect 52% of positive cases. Conclusion: The mgc2 real-time LAMP emerged as a more sensitive and accurate method for molecular detection of M. gallisepticum than RSA. Robustness and precision give it applicability as a potential field diagnostic tool for M. gallisepticum control. The study will be beneficial in reducing economic losses that M. gallisepticum inflicts on the poultry industry. This is the first reported development of a real-time LAMP assay based on the amplification of the mgc2 gene sequence using an ESEQuant tube scanner for galline M. gallisepticum detection. |
| Databáze: | OpenAIRE |
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