کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6115970 | 1215939 | 2013 | 4 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Quadraplex qRT-PCR assay for the simultaneous detection of Eastern equine encephalitis virus and West Nile virus
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
میکروبیولوژی و بیوتکنولوژی کاربردی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Quadraplex qRT-PCR assay for the simultaneous detection of Eastern equine encephalitis virus and West Nile virus Quadraplex qRT-PCR assay for the simultaneous detection of Eastern equine encephalitis virus and West Nile virus](/preview/png/6115970.png)
چکیده انگلیسی
In order to increase testing throughput and reduce cost, we developed a multiplex real-time assay that identifies both Eastern equine encephalitis virus and West Nile virus. The assay allows for the screening for the presence of both the nonstructural and envelope genes of both viruses simultaneously allowing for confirmatory testing to be done in a single assay. We utilized newly designed primers and probes, each labeled with a unique fluorescent label allowing for differentiation using an ABI 7500 real-time PCR machine. The use of Quanta Biosciences qScript XLT One-Step RT-qPCR® Toughmix allowed for a quadraplex assay without loss of sensitivity when compared to the previously run singleplex reaction as seen with viral RNA PFU control dilution series. There was no cross reactivity between the viruses within the reaction, and upon utilization of the assay during surveillance, there was no cross reactivity with other historically encountered arthropod-borne viruses. The results from the quantitative Reverse Transcriptase - Polymerase Chain Reaction were comparable to those achieved by cell culture which was performed on a subset of the field mosquito pools screened during the 2012 surveillance season. The multiplex assay resulted in savings in both time and resources for the lab and faster turn-around of results.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Diagnostic Microbiology and Infectious Disease - Volume 77, Issue 2, October 2013, Pages 129-132
Journal: Diagnostic Microbiology and Infectious Disease - Volume 77, Issue 2, October 2013, Pages 129-132
نویسندگان
Steven D. Zink, Susan A. Jones, Joseph G. Maffei, Laura D. Kramer,