کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
3408099 1593509 2007 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Detection and quantitation of Solenopsis invicta virus in fire ants by real-time PCR
موضوعات مرتبط
علوم زیستی و بیوفناوری ایمنی شناسی و میکروب شناسی ویروس شناسی
پیش نمایش صفحه اول مقاله
Detection and quantitation of Solenopsis invicta virus in fire ants by real-time PCR
چکیده انگلیسی

A quantitative real-time PCR (QPCR) method was developed to detect and quantify the amount of Solenopsis invicta virus (SINV) infecting individual ants of S. invicta. The two-step method utilized a gene-specific oligonucleotide primer targeting the SINV RNA-dependent RNA polymerase (RdRp) for cDNA synthesis. Dithiothreitol used in the cDNA synthesis step was found to significantly decrease the detection sensitivity for SINV RdRp and was therefore omitted. SINV RdRp cDNA was then quantified by QPCR using SYBR Green dye and a standard curve generated from SINV RdRp plasmid clones. A standard curve was successfully constructed from clones of the SINV RdRp region. A strong linear relationship [r2 = 0.998; y = (−3.63 ± 0.37)x + (39.19 ± 1.33)] between CT and starting SINV RdRp copy number was observed within a dynamic range of 5–5 × 106 copies. SINV RdRp copy number was determined with the optimized QPCR method in individual S. invicta ants taken from an infected field colony. Worker ants exhibited the highest RdRp copy number (2.1 × 109 copies/worker ant) and pupae exhibited the lowest (4.2 × 102 copies/pupa). Mean RdRp copy number was lowest in early larvae and pupae. Overall, SINV RdRp copy number increased through larval development, sharply declined during pupation, then sharply increased in adults.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 140, Issues 1–2, March 2007, Pages 132–139
نویسندگان
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