A Rapid and Simple Loop-Mediated Isothermal Amplification Assay for the Detection of Pseudomonas aeruginosa From Equine Genital Swabs

التفاصيل البيبلوغرافية
العنوان: A Rapid and Simple Loop-Mediated Isothermal Amplification Assay for the Detection of Pseudomonas aeruginosa From Equine Genital Swabs
المؤلفون: Noel Fitzpatrick, Jason Sawyer, Sarah North, Onyinye Diribe, Roberto M. La Ragione
المصدر: Journal of Equine Veterinary Science. 35:929-934
بيانات النشر: Elsevier BV, 2015.
سنة النشر: 2015
مصطلحات موضوعية: Equine, Pseudomonas aeruginosa, In silico, Loop-mediated isothermal amplification, Biology, Amplicon, biology.organism_classification, medicine.disease_cause, Rapid detection, Molecular biology, Microbiology, chemistry.chemical_compound, chemistry, medicine, Gene, Bacteria, DNA
الوصف: Pseudomonas aeruginosa is a causative agent of infectious reproductive disease in horses. Conventional methods for identification of this organism from equine genital swabs can be time consuming, laborious, and may rely on the presence of live bacteria. However, the use of loop-mediated isothermal amplification (LAMP) provides a simple and rapid detection method. We aimed to develop and evaluate a simple-to-use and interpret LAMP assay, based on the P. aeruginosa ecf X gene, capable of detecting P. aeruginosa from DNA extracted directly from equine genital swabs without culture or prior enrichment. The specificity of the assay was determined using DNA extracted from 29 different bacterial species, as well as in silico. The diagnostic sensitivity of the assay was determined through analysis of DNA extracted directly from 46 equine genital swabs. A P. aeruginosa –specific LAMP assay that could detect as few as 11 copies of its target in less than 15 minutes was developed. The reaction amplicon could be detected by real-time fluorescence or using a simple lateral flow device. The assay was specific to its target, showing no cross-reactivity or interference when tested against DNA extracted from 29 other bacterial species, as well as in silico. The diagnostic sensitivity of the assay was 86.8% (39 of 46) and 88.5% (40 of 46), when the amplicon was detected by real-time fluorescence and lateral flow device technology, respectively. The P. aeruginosa –specific LAMP assay provides a simple and rapid means of detecting the presence of viable and nonviable P. aeruginosa directly from equine genital swabs.
تدمد: 0737-0806
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::3958529e7d767460dd9956e01456ee68
https://doi.org/10.1016/j.jevs.2015.08.017
حقوق: CLOSED
رقم الأكسشن: edsair.doi...........3958529e7d767460dd9956e01456ee68
قاعدة البيانات: OpenAIRE