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对一只狗耳部分泌物中的印加蠕形螨进行聚合酶链反应扩增和DNA测序。

PCR amplification and DNA sequencing of Demodex injai from otic secretions of a dog.

作者信息

Milosevic Milivoj A, Frank Linda A, Brahmbhatt Rupal A, Kania Stephen A

机构信息

Department of Small Animal Clinical Sciences, University of Tennessee, Knoxville, TN 37996-4544, USA.

出版信息

Vet Dermatol. 2013 Apr;24(2):286-e66. doi: 10.1111/vde.12010. Epub 2013 Feb 18.

Abstract

BACKGROUND

The identification of Demodex mites from dogs is usually based on morphology and location. Mites with uncharacteristic features or from unusual locations, hosts or disease manifestations could represent new species not previously described; however, this is difficult to determine based on morphology alone.

HYPOTHESIS/OBJECTIVES: The goal of this study was to identify and confirm Demodex injai in association with otitis externa in a dog using PCR amplification and DNA sequencing.

METHODS

Otic samples were obtained from a beagle in which a long-bodied Demodex mite was identified. For comparison, Demodex mite samples were collected from a swab and scraping of the dorsal skin of a wire-haired fox terrier and an otic sample from a dog with generalized and otic demodicosis. To identify the Demodex mite, DNA was extracted, and 16S rRNA was amplified by PCR, sequenced and compared with Demodex sequences available in public databases and from separate samples morphologically diagnosed as D. injai and Demodex canis.

RESULTS

PCR amplification of the long-bodied mite rRNA DNA obtained from otic samples was approximately 330 bp and was identical to that from the mite morphologically identified as D. injai obtained from the dorsal skin of a dog. Furthermore, the examined mite did not have any significant homology to any of the reported genes from Demodex spp.

CONCLUSIONS

These results confirmed that the demodex mites in this case were D. injai.

摘要

背景

从犬只身上鉴定蠕形螨通常基于形态学和寄生部位。具有非典型特征或来自不寻常部位、宿主或疾病表现的螨可能代表以前未描述过的新物种;然而,仅根据形态学很难确定。

假设/目的:本研究的目的是通过聚合酶链反应(PCR)扩增和DNA测序,鉴定并确认一只患有外耳炎的犬只身上的印氏蠕形螨。

方法

从一只小猎犬的耳部获取样本,在该样本中鉴定出一种长体型的蠕形螨。为作比较,从一只刚毛猎狐梗的背部皮肤擦拭和刮片中收集蠕形螨样本,并从一只患有全身性和耳部蠕形螨病的犬只获取耳部样本。为鉴定蠕形螨,提取DNA,通过PCR扩增16S rRNA,进行测序,并与公共数据库中以及从形态学诊断为印氏蠕形螨和犬蠕形螨的单独样本中获得的蠕形螨序列进行比较。

结果

从耳部样本中获得的长体型螨rRNA DNA的PCR扩增产物约为330 bp,与从一只犬背部皮肤形态学鉴定为印氏蠕形螨的螨的扩增产物相同。此外,所检测的螨与已报道的任何蠕形螨属基因均无显著同源性。

结论

这些结果证实该病例中的蠕形螨为印氏蠕形螨。

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