PCR-RFLP method for the identification of four members of the Anopheles annularis group of mosquitoes (Diptera: Culicidae)
PCR-RFLP method for the identification of four members of the Anopheles annularis group of mosquitoes (Diptera: Culicidae)
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DOI：
10.1016/j.trstmh.2006.03.007
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发表时间：
2007-03-01
影响因子：
2.2
通讯作者：
Sharma, Yagya D.
中科院分区：
文献类型：
--
作者：
Alam, Mohammad Tauqeer;Das, Manoj K.;Sharma, Yagya D.
环按蚊广泛分布于东南亚，可能是当地重要的疟疾媒介。该类群的成员在形态上非常相似，通常很难区分，特别是 A. nivipes 和 A. philippinensis。我们报告了 A 组四个成员的 rDNA 内部转录间隔区 2 (ITS2) 和结构域 3 (D3) 区域的序列分析。 nivipes、A. philippinensis、A. anialis 和 A. pallidus - 及其分子鉴定方法。标本之间未检测到种内序列变异，而 ITS2 区域的种间序列差异大于 D3 区域。四个物种的D3序列的比较显示，在A. nivipes中存在2个SmaI限制性位点，但在A. philippinensis、A.环状和A. pallidus中只有1个位点。 Apal 位点存在于 A. philippinensis 和 A. pallidus 中，而 Ncol 位点仅存在于 A. pallidus 中。用 Smal、Apal 和 Ncol 分别限制性消化 D3 片段的 PCR 产物，产生了所有四个物种的独特模式。我们首次提出了一种 PCR-RFLP 方法来区分环环蚊类的四种成员。 (c) 2006 年皇家热带医学和卫生学会。由爱思唯尔有限公司出版。保留所有权利。
The Anopheles annularis group of mosquitoes is widely distributed in Southeast Asia and may be locally important as malaria vectors. Members of this group are morphologically very similar and often difficult to distinguish, particularly A. nivipes and A. philippinensis. We report the sequence analysis of the rDNA internal transcribed spacer 2 (ITS2) and Domain-3 (D3) regions of the four members of the A. annularis group-A. nivipes, A. philippinensis, A. annularis and A. pallidus-and a method for their molecular identification. No intraspecies sequence variation was detected among the specimens, while interspecific sequence differences were greater for ITS2 than the D3 regions. Comparison of the D3 sequences of the four species revealed two Smal restriction sites in A. nivipes, but only one site in A. philippinensis, A. annularis and A. pallidus. The Apal site was present in both A. philippinensis and A. pallidus, while an Ncol site was present in A. pallidus only. Restriction digestion of the PCR products of D3 fragment individually with Smal, Apal and Ncol produced a distinctive pattern for all the four species. We present, for the first time, a PCR-RFLP method to distinguish the four members of the A. annularis group of mosquitoes. (c) 2006 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.