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1、VIROLOGICASINICA2016,31(1):78–80DOI:10.1007/s12250-015-3703-3LETTERLongitudinalsurveillanceofSARS-likecoronavirusesinbatsbyquantitativereal-timePCRDearEditor,are expressed as genome copies per gram of bat feces(copies/g).The 2002–2003 severe acute respiratory syndromeThe qRT-PCRs were perfo
2、rmed using an AgPath-coronavirus (SARS-CoV) (Drosten et al., 2003) causedID™ One-Step RT-PCR Kit (Applied Biosystems) ac-human pandemics that began in China and spread glob-cording to the manufacturer’s instructions. Each 25-μLally. Subsequently, diverse SARS-like coronavirusesreaction mixt
3、ure contained 12.5 μL of 2× RT-PCR buf-(SL-CoVs) have been detected in horseshoe bats infer, 1 μL of RT-PCR Enzyme Mix, 400 nmol/L eachChina, Europe, and Africa (Li et al., 2005; Tong et al.,primer, 120 nmol/L probe primer, and 1 μL of nucleic2009; Drexler et al., 2010). Recently, we found
4、SL-CoVsacid extract. Amplification was carried out in 96-wellwith high genetic diversity in a single bat colony pre-plates using the StepOne PCR system (Applied Biosys-dominantly roosted by Chinese horseshoe bats (Rhinolo-tems). Thermocycling conditions were as follows: 10phussinicus) in Ku
5、nming, Yunnan province (Ge et al.,min at 45 °C for reverse transcription, 10 min at 95 °C2013). Two of these SL-CoVs are able to use humanfor activation of the Taq DNA polymerase, and 40 cyclesACE2 as a receptor for cell entry (Ge et al., 2013; Men-of 95 °C for 15 sec and 60 °C for 45 sec.
6、Each run in-achery et al., 2015), highlighting the risk of this group ofcluded three viral positive control templates and one neg-viruses to humans and the importance of long-term sur-ative control to evaluate assay performance. A positiveveillance.result was defined as a well-defined expon
7、ential fluores-We conducted a longitudinal surveillance study of batcence curve that crossed the cycle threshold (Ct) withinSL-CoVs using quantitative real-time PCR (qRT-PCR)38 cycles. A specimen with a Ctvalue >36 was assayedtargeting the nucleocapsid (
