绵羊梨型虫PCR方法的建立及序列分析
发布时间：2018-09-07 12:49
【摘要】：本研究建立了绵羊梨型虫的检测方法,并对退火温度和循环次数进行梯度优化。选取条带最亮且无杂带的温度和循环次数作为PCR检测方法的优化结果,将新建的PCR方法与传统的吉姆萨染色法结果进行对比,将阳性的PCR产物进行测序。结果表明:PCR反应的最佳体系为94℃预变性5 min;94℃变性1 min,56℃退火90s,72℃延伸1min,共循环39次;72℃延伸10min,4℃保存;在50份临床样品的检测中,PCR法检出的阳性样品有13份,阳性率为26%(13/50),吉姆萨染色发检出的阳性样品有8份,阳性率为16%(8/50),由PCR方法检测出的阳性样品与吉姆萨染色发检出的阳性样品符合率达100%。阳性PCR产物的测序结果的同源性分别为100%和98%。本研究证实PCR法的灵敏度和准确性高于吉姆萨氏染色镜检法。
[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，
本文编号：2228314
[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，
本文编号：2228314
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