CAJ | 학술논문

目的：建立鼠痘病毒的荧光定量Taqman-PCR检测方法，以期能快速、准确、灵敏、特异的检出鼠痘病毒。方法经过比对和筛选，本研究选取鼠痘病毒的ERPV＿027基因480－800位序列段，作为引物设计位点设计引物和探针，并对该引物对和探针的特异性、灵敏性、稳定性和重复性进行检测。结果本研究建立的方法，对鼠痘病毒的检测极限是68 copies／μL；该方法特异性强，只对鼠痘病毒特定片段进行特异性扩增，而对小鼠肝炎病毒、仙台病毒、沙门氏菌等其它病毒、细菌无扩增；该方法稳定性和重复性均较好。结论本研究成功建立了检测鼠痘病毒的荧光定量Taqman-PCR方法，该方法特异性强、灵敏度高，且具有较好的稳定性和重复性，具有广阔的应用价值。
목적：건립서두병독적형광정량Taqman-PCR검측방법，이기능쾌속、준학、령민、특이적검출서두병독。방법경과비대화사선，본연구선취서두병독적ERPV＿027기인480－800위서렬단，작위인물설계위점설계인물화탐침，병대해인물대화탐침적특이성、령민성、은정성화중복성진행검측。결과본연구건립적방법，대서두병독적검측겁한시68 copies／μL；해방법특이성강，지대서두병독특정편단진행특이성확증，이대소서간염병독、선태병독、사문씨균등기타병독、세균무확증；해방법은정성화중복성균교호。결론본연구성공건립료검측서두병독적형광정량Taqman-PCR방법，해방법특이성강、령민도고，차구유교호적은정성화중복성，구유엄활적응용개치。
Objective To establish a fluorescence quantitative Taqman-PCR method for rapid and accurate detection of mouse poxvirus.Methods After sequence alignment and comparison, ERPV_027 gene was selected as the primer and probe design gene.Furthermore, the specificity, sensitivity, stability and reproducibility of these primers and probes were detected.Results The detection limitation of this method was 68 copies/μL.Data showed that this method has high specificity, which specifically amplifies mouse poxvirus, with no amplification signal of mouse hepatitis virus, Sendai virus, Salmonella and some other viruses and bacteria.This method also showed good stability and reproducibility. Conclusions This study has successfully established a fluorescence quantitative Taqman-PCR method for detection of mouse poxvirus, with high specificity, sensitivity, good stability and reproducibility, and a broad application potential.