本文關(guān)鍵詞： 豬繁殖與呼吸綜合征病毒 不對(duì)稱PCR HRM　出處：《中國獸醫(yī)科學(xué)》2017年12期 　論文類型：期刊論文

【摘要】：為建立一種快速鑒別豬繁殖與呼吸綜合征病毒(PRRSV)疫苗株GDr180株與其他毒株的PCR-HRM檢測(cè)方法,比對(duì)NCBI登錄的PRRSV全基因序列,篩選GDr180株與其他株的差異位點(diǎn)。PCR反應(yīng)體系在加入LC Green染料及3′端封閉的探針下進(jìn)行不對(duì)稱PCR擴(kuò)增,在HRM功能模式下進(jìn)行PCR產(chǎn)物的熔解曲線分析。結(jié)果顯示,GDr180株P(guān)CR產(chǎn)物的熔解曲線探針峰Tm值為72.64±0.39℃,而PRRSV其他株的探針峰Tm值為65.63±0.69℃或無探針峰,兩者差異極顯著。用本方法引物擴(kuò)增非PRRSV如豬流行性腹瀉病毒(PEDV)、偽狂犬病病毒(PRV)、豬瘟病毒(CSFV)、豬圓環(huán)病毒(PCV)和豬輪狀病毒(Po RV)無特異性目的條帶,標(biāo)準(zhǔn)品檢測(cè)靈敏度為1 000 copies/L。本研究建立的PCR-HRM是一種高通量、經(jīng)濟(jì)、簡便和可靠的用于鑒別PRRSV GDr180株與其他毒株的方法。
[Abstract]:In order to establish a PCR-HRM method for rapid identification of porcine reproductive and respiratory syndrome virus (PRRSVV) vaccine strains from other strains, the whole PRRSV gene sequence registered by NCBI was compared. Screening the difference sites between GDr180 strain and other strains. PCR reaction system was amplified by asymmetric PCR with LC Green dye and 3 'closed probe. The melting curves of PCR products were analyzed in HRM functional mode. The results showed that the probe peak TM value of PCR products was 72.64 鹵0.39 鈩，

本文編號(hào)：1497719