کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6134853 | 1593490 | 2011 | 7 صفحه PDF | دانلود رایگان |
The development of a rapid and sensitive system for detecting influenza viruses is a high priority for controlling future epidemics and pandemics. Quantitative real-time PCR is often used for detecting various kinds of viruses; however, it requires more than 2Â h per run. Detection assays were performed with super high-speed RT-PCR (SHRT-PCR) developed according to a newly designed heating system. The new method uses a high-speed reaction (18Â s/cycle; 40 cycles in less than 20Â min) for typing influenza viruses. The detection limit of SHRT-PCR was 1 copy/reaction and 10â1 plaque-forming unit/reaction for viruses in culture supernatants during 20Â min. Using SHRT-PCR, 86 strains of influenza viruses isolated by the Tokyo Metropolitan Institute of Public Health were tested; the results showed 100% sensitivity and specificity for each influenza A and B virus, and swine-origin influenza virus. Twenty-seven swabs collected from the pharyngeal mucosa of outpatients were also tested, showing positive signs for influenza virus on an immunochromatographic assay; the results between SHRT-PCR and immunochromatography exhibited 100% agreement for both positive and negative results. The rapid reaction time and high sensitivity of SHRT-PCR makes this technique well suited for monitoring epidemics and pre-pandemic influenza outbreaks.
⺠A super high speed qRT-PCR (SHRT-PCR) was developed. RT reaction and PCR (40 cycles) can be completed in less than 20 min. ⺠Despite the high speed, the sensitivity and specificity of the SHRT-PCR are comparable to conventional qRT-PCR system. ⺠The SHRT-PCR can be used to test clinical samples.
Journal: Journal of Virological Methods - Volume 178, Issues 1â2, December 2011, Pages 75-81