Università degli Studi di Urbino Carlo Bo, Dipartimento di Scienze Biomolecolari, Via A. Saffi, 2 Urbino, 61029, Italy.
Vet Res Commun. 2009 Oct;33(7):611-8. doi: 10.1007/s11259-009-9210-y. Epub 2009 Feb 13.
In the present work mites previously identified as Dermanyssus gallinae De Geer (Acari, Mesostigmata) using morphological keys were investigated by molecular tools. The complete internal transcribed spacer 1 (ITS1), 5.8S ribosomal DNA, and ITS2 region of the ribosomal DNA from mites were amplified and sequenced to examine the level of sequence variations and to explore the feasibility of using this region in the identification of this mite. Conserved primers located at the 3'end of 18S and at the 5'start of 28S rRNA genes were used first, and amplified fragments were sequenced. Sequence analyses showed no variation in 5.8S and ITS2 region while slight intraspecific variations involving substitutions as well as deletions concentrated in the ITS1 region. Based on the sequence analyses a nested PCR of the ITS2 region followed by RFLP analyses has been set up in the attempt to provide a rapid molecular diagnostic tool of D. gallinae.
在本工作中，我们使用分子工具研究了先前通过形态学特征鉴定为鸡皮刺螨（蜱螨目，中气门目）的螨类。从螨类中扩增并测序了核糖体 DNA 的完整内部转录间隔区 1（ITS1）、5.8S 核糖体 DNA 和 ITS2 区，以检查序列变异水平，并探讨该区域在鉴定这种螨类中的可行性。首先使用位于 18S 末端 3'端和 28S rRNA 基因 5'端的保守引物进行扩增，然后对扩增片段进行测序。序列分析显示 5.8S 和 ITS2 区没有变异，而在 ITS1 区存在轻微的种内变异，包括替换以及集中的缺失。基于序列分析，我们建立了 ITS2 区的嵌套 PCR 及其随后的 RFLP 分析，试图提供一种快速的鸡皮刺螨分子诊断工具。
