Brzo i vidljivo petljom posredovano izotermno umnažanje za dokazivanje Brucella spp. i njegova primjena u epidemiologiji bruceloze goveda
العنوان: | Brzo i vidljivo petljom posredovano izotermno umnažanje za dokazivanje Brucella spp. i njegova primjena u epidemiologiji bruceloze goveda |
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المؤلفون: | Kumaragurubaran Karthik, Rajesh Rathore, Prasad Thomas, Konasagara N. Viswas, Rajesh K. Agarwal, Valsala Rekha, Ramesh V. Jagapur, Kuldeep Dhama |
المصدر: | Veterinarski arhiv Volume 86 Issue 1 |
بيانات النشر: | Faculty of Veterinary Medicine University of Zagreb, 2016. |
سنة النشر: | 2016 |
مصطلحات موضوعية: | genetic structures, petljom posredovano izotermno umnažanje, brucele, početnice, omp25, SYBR zeleno, eye diseases, visual loop mediated isothermal amplification test, Brucella, loop primers, SYBR green |
الوصف: | Brucellosis is a devastating disease, once an abortion storm starts in a herd, and hence early diagnosis is important. Loop mediated isothermal amplification (LAMP) has enlightened the darker part of the diagnostic world by its simplicity and swiftness, and has the advantage of working in isothermal conditions so that it can be applied to the field. LAMP for Brucella spp., that already exists has not been exploited for visual detection and the time needed for completion of the reaction has not been reduced. In the present study, an attempt was made to seal these cracks by developing a fast and visually detectable LAMP for Brucella spp., which needs only 30 min detection time, and SYBR green aided easier result visualization. The LAMP test targeting the omp25 gene was found to be highly sensitive and specific for detecting Brucella spp. Comparison of the LAMP test with the available PCR assay revealed that the LAMP test was more sensitive than PCR, following testing of 438 field samples of cattle origin, and it showed prevalence of Brucella of 14.7 % in different parts of India. Being highly sensitive, specific and speedy, the standardized visual LAMP test can be widely used for epidemiological surveys of this economically important and zoonotic pathogen. Bruceloza je razorna bolest kad se u stadu pojavi u obliku pobačaja što iziskuje brzo postavljanje dijagnoze. Upotreba metode nazvane „petljom posredovano izotermno umnažanje“ (engl. loop mediated isothermal amplification, LAMP)“ omogućila je poboljšanje dijagnostike svojom jednostavnošću i brzinom, a prednost joj je da se može izvesti na postojanoj temperaturi pa se može primijeniti u terenskim uvjetima. U dosada razvijenim postupcima LAMP-a nije bio rabljen vizualni dokaz i nije se uspjelo skratiti vrijeme reakcije. U ovom istraživanju pokušalo se ukloniti te nedostatke razvitkom brzog postupka za vizualni dokaz Brucella spp., za što je potrebno samo 30 minuta i SYBR zeleno. LAMP test za dokaz gena omp25 pokazao se visoko osjetljivim i specifičnim za dokaz Brucella spp. Pretraga 438 terenskih uzoraka podrijetlom od goveda pokazala je da je LAMP osjetljiviji od raspoloživih testova PCR-a. Tim je testom dokazana prevalencija bruceloze od 14,7 % u različitim dijelovima Indije. Kao vrlo osjetljiv, specifičan i brz, LAMP test se može naširoko upotrijebiti u epidemiološkim istraživanjima bruceloze. |
وصف الملف: | application/pdf |
اللغة: | English |
تدمد: | 1331-8055 0372-5480 |
URL الوصول: | https://explore.openaire.eu/search/publication?articleId=od_______951::07ba98720994f30165139522066ac64c https://hrcak.srce.hr/file/226131 |
حقوق: | OPEN |
رقم الأكسشن: | edsair.od.......951..07ba98720994f30165139522066ac64c |
قاعدة البيانات: | OpenAIRE |
تدمد: | 13318055 03725480 |
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