کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6132802 | 1593443 | 2016 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Development and evaluation of a reverse transcription-insulated isothermal polymerase chain reaction (RT-iiPCR) assay for detection of equine arteritis virus in equine semen and tissue samples using the POCKIT⢠system
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کلمات کلیدی
EVAVNTEHVOIEEAVMGBEMEMVBSRT-qPCRFAMORF6-carboxyfluorescein - 6-کربوکسی فلوئورسینS/N - S / NEquine viral arteritis - آرتریت ویروسی اسبVirus neutralization test - آزمون خنثی سازی ویروسEagle’s Minimum Essential Medium - حداقل ضروری متوسط عقابReal-time RT-PCR - زمان واقعی RT-PCRWorld Organisation for Animal Health - سازمان جهانی بهداشت حیواناتAbortion - سقط جنینopen reading frame - قاب خواندن بازSignal-to-noise ratio - نسبت سیگنال به نویزplaque-forming unit - واحد پلاک سازیEquine arteritis virus - ویروس آرتروز اسبEquine herpesvirus - ویروس هرپس اسبpfu - پفو
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Equine arteritis virus (EAV) is the causative agent of equine viral arteritis (EVA), a respiratory and reproductive disease of horses. Most importantly, EAV induces abortion in pregnant mares and can establish persistent infection in up to 10-70% of the infected stallions, which will continue to shed the virus in their semen. The objective of this study was to develop and evaluate a reverse transcription insulated isothermal polymerase chain reaction (RT-iiPCR) for the detection of EAV in semen and tissue samples. The newly developed assay had a limit of detection of 10 RNA copies and a 10-fold higher sensitivity than a previously described real-time RT-PCR (RT-qPCR). Evaluation of 125 semen samples revealed a sensitivity and specificity of 98.46% and 100.00%, respectively for the RT-qPCR assay, and 100.00% and 98.33%, respectively for the RT-iiPCR assay. Both assays had the same accuracy (99.2%, k = 0.98) compared to virus isolation. Corresponding values derived from testing various tissue samples (n = 122) collected from aborted fetuses, foals, and EAV carrier stallions are as follows: relative sensitivity, specificity, and accuracy of 88.14%, 96.83%, and 92.62% (k = 0.85), respectively for the RT-qPCR assay, and 98.31%, 92.06%, and 95.08% (k = 0.90), respectively for the RT-iiPCR assay. These results indicate that RT-iiPCR is a sensitive, specific, and a robust test enabling detection of EAV in semen and tissue samples with very considerable accuracy. Even though the RT-qPCR assay showed a sensitivity and specificity equal to virus isolation for semen samples, its diagnostic performance was somewhat limited for tissue samples. Thus, this new RT-iiPCR could be considered as an alternative tool in the implementation of EAV control and prevention strategies.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 234, August 2016, Pages 7-15
Journal: Journal of Virological Methods - Volume 234, August 2016, Pages 7-15
نویسندگان
Mariano Carossino, Pei-Yu A. Lee, Bora Nam, Ashley Skillman, Kathleen M. Shuck, Peter J. Timoney, Yun-Long Tsai, Li-Juan Ma, Hsiao-Fen G. Chang, Hwa-Tang T. Wang, Udeni B.R. Balasuriya,