【摘要】：本研究建立了綿羊梨型蟲的檢測方法,并對退火溫度和循環(huán)次數進行梯度優(yōu)化。選取條帶最亮且無雜帶的溫度和循環(huán)次數作為PCR檢測方法的優(yōu)化結果,將新建的PCR方法與傳統(tǒng)的吉姆薩染色法結果進行對比,將陽性的PCR產物進行測序。結果表明:PCR反應的最佳體系為94℃預變性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100%。陽性PCR產物的測序結果的同源性分別為100%和98%。本研究證實PCR法的靈敏度和準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：2228314