کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10030476 | 1593416 | 2018 | 28 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Development of duplex fluorescence-based loop-mediated isothermal amplification assay for detection of Mycoplasma bovis and bovine herpes virus 1
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کلمات کلیدی
BHV-1SRSVPPRVOIEBRVRPVBTVFMDVVSVCVCCIACUCETECBVDVBRDPeste des petits ruminants virus - Peste des petits معتادین ویروسEscherichia coli - اشریشیا کُلیBovine respiratory disease - بیماری تنفسی گاوFluorescent detection - تشخیص فلورسنتBovine rotavirus - روتاویروس گاوWorld Organisation for Animal Health - سازمان جهانی بهداشت حیواناتInstitutional Animal Care and Use Committee - سازمان مراقبت و مراقبت از حیواناتMycoplasma bovis - مایکوپلاسما bovisBluetongue virus - ویروس BluetongueVesicular stomatitis virus - ویروس vesicular stomatitisFoot-and-mouth disease virus - ویروس آنفلوآنزای مرغیBovine virus diarrhoea virus - ویروس اسهال ویروس گاوBovine herpes virus 1 - ویروس تبخال تبخال 1bovine respiratory syncytial virus - ویروس سونسیتیال تنفسی گاوRinderpest virus - ویروس گوسفند
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Mycoplasma bovis (MB) and bovine herpes virus 1 (BHV-1) are two important pathogens that cause bovine respiratory disease in the beef feedlot and dairy industries. The aim of this study was to develop and validate a duplex fluorescence-based loop-mediated isothermal amplification (DLAMP) assay for simultaneous detection of MB and BHV-1. Two sets of specific primers for each pathogen were designed to target the unique sequences of the MB uvrC gene and the BHV-1 gB gene. The inner primer for BHV-1 was synthesized with the fluorophore FAM at the 5â² end to detect the BHV-1 gB gene, and the inner primer for MB was synthesized with the fluorophore CY5 at the 5â² end to detect the MB uvrC gene. The DLAMP reaction conditions were optimized for rapid and specific detection of MB and BHV-1. The DLAMP assay developed here could specifically detect MB and BHV-1 without cross-reaction with other known non-target bovine pathogens. The sensitivity of this DLAMP assay was as low as 2âÃâ102 copies for recombinant plasmids containing the MB and BHV-1 target genes. In a detection test of 125 clinical samples, the positive rates for MB, BHV-1 and co-infection were 44.8%, 13.6% and 1.6%, respectively. Furthermore, the sensitivity and specificity of DLAMP were determined as 95%-96.6% and 100%, respectively, of those of field sample detection by the real-time polymerase chain reaction (PCR) assay recommended by the World Organisation for Animal Health. Overall, DLAMP provides a rapid, sensitive and specific assay for the identification of MB and BHV-1 in clinical specimens and for epidemiological surveillance.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 261, November 2018, Pages 132-138
Journal: Journal of Virological Methods - Volume 261, November 2018, Pages 132-138
نویسندگان
Qing Fan, Zhixun Xie, Zhiqin Xie, Liji Xie, Jiaoling Huang, Yanfan Zhang, Tingting Zeng, Minxiu Zhang, Sheng Wan, Sisi Luo, Jiabo Liu, Xianwen Deng,