کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6132969 | 1593450 | 2016 | 37 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A new method for simultaneous detection and discrimination of Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) using real time PCR with high resolution melting (HRM) analysis
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کلمات کلیدی
DPIHRMBoHV-1MDBKBoHV-5Bovine herpesvirus type 5Real time PCR - PCR در زمان واقعیcycle threshold - آستانه چرخهMolecular detection - تشخیص مولکولیMelting Temperature - دمای ذوبHigh resolution melting - ذوب شدن با وضوح بالاdays post-infection - روز بعد از عفونتbovine herpesvirus - ویروس گاوBovine herpesvirus type 1 - ویروس گاو نوع 1
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are antigenically and genetically similar. The aim of this study was to develop a simple and reliable one-step real time PCR assay with high resolution melting (HRM) analysis for the simultaneous detection and differentiation of BoHV-1 and BoHV-5. Optimization of assay conditions was performed with DNA from reference strains. Then, DNA from field isolates, clinical samples and tissue samples of experimentally infected animals were studied by real time PCR-HRM. An efficient amplification of real time PCR products was obtained, and a clear melting curve and appropriate melting peaks for both viruses were achieved in the HRM curve analysis for BoHV type identification. BoHV was identified in all of the isolates and clinical samples, and BoHV types were properly differentiated. Furthermore, viral DNA was detected in 12/18 and 7/18 samples from BoHV-1- and BoHV-5-infected calves, respectively. Real time PCR-HRM achieved a higher sensitivity compared with virus isolation or conventional PCR. In this study, HRM was used as a novel procedure. This method provides rapid, sensitive, specific and simultaneous detection of bovine alpha-herpesviruses DNA. Thus, this technique is an excellent tool for diagnosis, research and epidemiological studies of these viruses in cattle.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 227, January 2016, Pages 14-22
Journal: Journal of Virological Methods - Volume 227, January 2016, Pages 14-22
نویسندگان
M.S. Marin, S. Quintana, M.R. Leunda, M. Recavarren, I. Pagnuco, E. Späth, S. Pérez, A. Odeón,