一种犬耳痒螨PCR检测方法的建立及应用  

Establishment of PCR Method for the Detection of Otodectes cynotis and Its Application

作　　者：杜嘉玥 缑静敏 要慧中 梁高星 林青[1] DU Jia-yue;GOU Jing-min;YAO Hui-zhong;LIANG Gao-xing;LIN Qing(College of Veterinary Medicine,Northwest A&F University,Yangling,712100,China;Xi’an Veterinary Teaching Hospital,Northwest A&F University,Xi’an,710065,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]西北农林科技大学西安动物医院,陕西西安710068

出　　处：《动物医学进展》2023年第5期27-33,共7页Progress In Veterinary Medicine

基　　金：西安西北农林科大动物医院临床科研基金资助项目(K4040121223);陕西诺威利华生物科技有限公司协作项目(K4040121325)。

摘　　要：为了能从犬、猫耳垢中对犬耳痒螨(Otodectes cynotis)进行特异性检测,基于犬耳痒螨18S-ITS1基因序列设计特异性引物,通过对退火温度等条件进行优化,建立犬耳痒螨特异性PCR检测方法。结果显示,该方法能成功从犬耳痒螨DNA样本中扩增出352 bp的特异性条带,特异性良好;其最低检测DNA浓度为0.897 fg/μL,敏感性较高。用所建立的PCR检测方法对78只犬、猫的耳垢样本进行检测,犬耳痒螨的阳性率为66.7%,高于显微镜检测结果(阳性率为62.8%);对52份经PCR方法检测的犬耳痒螨阳性样本进行种群结构分析,未发现犬耳痒螨rDNA ITS1序列的遗传变异出现地理差异和宿主特异情况。结果表明,建立的PCR方法能够应用于临床检测犬、猫耳垢样本中的犬耳痒螨,为犬、猫耳螨病的诊断提供了一种辅助方法。In order to detect Otodectes cynotis specifically from earwax in dogs and cats,the PCR method for the detection of O.cynotis was established by the design of specific primers on the basis of 18S-ITS1 gene sequence through optimizing annealing temperature and other conditions in this study.The results showed that the method had good sensitivity,which could successfully amplified a specific fragment of about 352 bp from positive samples of O.cynotis.The minimum detection concentration was 0.897 fg/μL,indicating high sensitivity.The PCR method was used to detect the earwax samples from 78 dogs and cats.Results revealed that the positive rate of O.cynotis was 66.7%,which was higher than using microscopy(62.8%).The population structure of 52 positive samples of O.cynotis was analyzed,and no geographical differences and host-specific characteristics were found in genetic variation of rDNA ITS1 sequence of O.cynotis.The results reveal that the PCR method established in this study can be applied to the clinical detection of O.cynotis in the earwax of dogs and cats,which can provide an auxiliary method for the diagnosis of otodectosis in dogs and cats.

关 键 词：犬耳痒螨 犬 猫 聚合酶链反应 

分 类 号：S855.9[农业科学—临床兽医学] S858.292[农业科学—兽医学]