【摘要】：本研究建立了綿羊梨型蟲的檢測(cè)方法,并對(duì)退火溫度和循環(huán)次數(shù)進(jìn)行梯度優(yōu)化。選取條帶最亮且無雜帶的溫度和循環(huán)次數(shù)作為PCR檢測(cè)方法的優(yōu)化結(jié)果,將新建的PCR方法與傳統(tǒng)的吉姆薩染色法結(jié)果進(jìn)行對(duì)比,將陽性的PCR產(chǎn)物進(jìn)行測(cè)序。結(jié)果表明:PCR反應(yīng)的最佳體系為94℃預(yù)變性5 min;94℃變性1 min,56℃退火90s,72℃延伸1min,共循環(huán)39次;72℃延伸10min,4℃保存;在50份臨床樣品的檢測(cè)中,PCR法檢出的陽性樣品有13份,陽性率為26%(13/50),吉姆薩染色發(fā)檢出的陽性樣品有8份,陽性率為16%(8/50),由PCR方法檢測(cè)出的陽性樣品與吉姆薩染色發(fā)檢出的陽性樣品符合率達(dá)100%。陽性PCR產(chǎn)物的測(cè)序結(jié)果的同源性分別為100%和98%。本研究證實(shí)PCR法的靈敏度和準(zhǔn)確性高于吉姆薩氏染色鏡檢法。
[Abstract]:In this study, a method for detecting pears of sheep was established, and the annealing temperature and cycle times were optimized by gradient. The temperature and cycle times of the brightest and non-heterogenous bands were selected as the optimized results of PCR detection. The new PCR method was compared with the traditional Gimsa staining method and the positive PCR products were sequenced. The results showed that the optimal reaction system was 94 鈩，

本文編號(hào)：2228314