骆驼斯氏副柔线虫病流行病学调查及生前PCR诊断方法建立

发布时间：2018-04-24 03:30

本文选题：骆驼斯氏副柔线虫病 + 流行病学调查　；参考：《内蒙古农业大学》2015年硕士论文

【摘要】：斯氏副柔线虫病是严重危害骆驼养殖业的一种寄生虫病。为了解近几年内蒙古地区骆驼斯氏副柔线虫病的流行情况,建立一种骆驼斯氏副柔线虫病的生前诊断方法。本试验于2013-2015年每年冬季,对内蒙古地区骆驼主产地的骆驼斯氏副柔线虫病流行情况进行了调查。利用斯氏副柔线虫rDNA ITS基因高度特异区域设计斯氏副柔线虫特异性引物,通过7种骆驼常见的消化道线虫及斯氏副柔线虫虫卵,对所设计引物的特异性、敏感性进行了验证,并采用3种方法对人工模拟阳性骆驼粪便中斯氏副柔线虫虫卵DNA进行了提取,以研究骆驼斯氏副柔线虫病生前诊断方法。结果显示,内蒙古地区四个骆驼主产地的108峰骆驼斯氏副柔线虫平均感染率为81.3%,最高感染强度为1272条/峰,雌雄虫比例为1.7：1-2.4：1,被调查的骆驼中巴彦淖尔市感染率为100%,乌兰察布市为85%,阿拉善盟为70.9%,锡林郭勒盟为57.1%。试验所设计的引物特异性强、敏感性高,目的片段为551bp,可检出15枚斯氏副柔线虫虫卵。改良CTAB+SiO2吸附法提取人工模拟阳性骆驼粪便中斯氏副柔线虫虫卵DNA,虫卵检出率为80%,优于试剂盒提取法及传统饱和酚-氯仿抽提法。本试验结果表明,近年来内蒙古地区骆驼斯氏副柔线虫病流行依然普遍,所设计的斯氏副柔线虫特异性引物及筛选出的DNA提取方法可用于骆驼斯氏副柔线虫病阳性粪便检测,为骆驼斯氏副柔线虫病生前诊断及流行病学调查奠定了基础。
[Abstract]:Nematode nematode disease is a parasitic disease that seriously endangers camel farming. In order to understand the prevalence of nematode in camel in Inner Mongolia in recent years, an antemortem diagnostic method was established. In the winter of 2013-2015, a survey was conducted on the prevalence of nematode nematodes in the main producing areas of camels in Inner Mongolia. Using the highly specific region of rDNA ITS gene, the specific primers were designed. The specificity and sensitivity of the primers were verified by seven common alimentary canal nematodes and eggs of Camel. Three methods were used to extract the DNA from the feces of artificially simulated positive camels, and to study the methods of diagnosis of nematodes spp. The results showed that the average infection rate was 81.3 and the highest infection intensity was 1272 per peak. The infection rate of the camels investigated was 100 in Bayannur, 85in in Ulaanchab, 70.9 in Alashan and 57.1 in Xilingol. The primers designed were highly specific and sensitive, and the target fragment was 551bp.The 15 eggs of nematode skrjabini could be detected. The modified CTAB SiO2 adsorption method was used to extract the egg DNA from simulated positive camel feces. The detection rate of egg was 80%, which was superior to the extraction method of kit and traditional saturated phenol-chloroform extraction method. The results showed that the prevalence of nematode spp. In recent years was still widespread in Inner Mongolia. The designed primers and the DNA extraction method could be used to detect the feces of the positive cases of nematode camels. The results laid a foundation for the antemortem diagnosis and epidemiological investigation of Camel's nematode parasiliasis.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.24

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