Identification of the main malaria vectors in the Anopheles gambiae species complex using a TaqMan real-time PCR assay.
Identification of the main malaria vectors in the Anopheles gambiae species complex using a TaqMan real-time PCR assay.
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DOI：
10.1186/1475-2875-6-155
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发表时间：
2007-11-22
期刊：
影响因子：
3
通讯作者：
Field, Linda M.
中科院分区：
文献类型：
--
作者：
Bass, Chris;Williamson, Martin S.;Wilding, Craig S.;Donnelly, Martin J.;Field, Linda M.
冈比亚按蚊物种复合体由七种形态难以区分的兄弟蚊子组成。两种主要传疟媒介阿拉伯按蚊和按蚊的快速鉴定。严格意义上的冈比亚按蚊来自非媒介物种四环按蚊，通常需要将其作为媒介控制方案的一部分。目前最广泛使用的物种鉴定方法是针对核糖体DNA序列中物种特异性差异的多重PCR方法。虽然这种检测在许多研究中被证明是相当可靠的，但在聚合酶链式反应后需要额外的步骤，这使得它很耗时。最近，一种基于TaqMan单核苷酸多态基因分型的高通量检测方法可以检测和区分An。Gbiae S.S.和An.已有报道称阿拉比星。开发了一种新的TaqMan检测方法，用于区分主要的疟疾媒介(An.Arabiens和An.冈比亚亚种(Gbiae S.S.)和非向量An。在发现现有的TaqMan试验错误地鉴定了一种四环线虫之后。Quriannulatus，An.Merus和An.梅拉斯作为一名。冈比亚亚种在对来自撒哈拉以南非洲13个国家的450多个样本进行的盲目物种鉴定试验中，将这种新的TaqMan检测方法与现有的TaqMan检测方法和标准的PCR方法进行了比较。标准的PCR方法被发现具有较低的错误分数(<；1%)的特异性，然而，与TaqMan方法相比，它显示出明显更高的失败反应次数(15%)。新的媒介特异性TaqMan试验和现有的TaqMan试验都显示错误识别样本(0和0.54%)和失败反应(1.25%和2.96%)的数量非常少。在分析灵敏度测试中，新的TaqMan检测方法显示出非常低的检测阈值，因此可以在新鲜或硅胶干燥的蚊子的单条腿上使用，而不需要首先提取DNA。这项研究描述了一种快速而灵敏的检测方法，它非常有效地识别了蚊子的两种主要疟疾媒介。冈比亚亚科与非媒介近缘种的复合体。该方法基于TaqMan SNP基因分型，可用于从干燥标本中筛选单腿。在一些地区，一个。MERUS/MLAS/bwambae是分布受限的载体，不存在，它可以单独用于区分载体和非载体，也可以与Walker TaqMan试验结合用于区分An。Arabiens和An.冈比亚亚种
The Anopheles gambiae sensu lato species complex comprises seven sibling species of mosquitoes that are morphologically indistinguishable. Rapid identification of the two main species which vector malaria, Anopheles arabiensis and An. gambiae sensu stricto, from the non-vector species Anopheles quadriannulatus is often required as part of vector control programmes. Currently the most widely used method for species identification is a multiplex PCR protocol that targets species specific differences in ribosomal DNA sequences. While this assay has proved to be reasonably robust in many studies, additional steps are required post-PCR making it time consuming. Recently, a high-throughput assay based on TaqMan single nucleotide polymorphism genotyping that detects and discriminates An. gambiae s.s and An. arabiensis has been reported. A new TaqMan assay was developed that distinguishes between the main malaria vectors (An. arabiensis and An. gambiae s.s.) and the non-vector An. quadriannulatus after it was found that the existing TaqMan assay incorrectly identified An. quadriannulatus, An. merus and An. melas as An. gambiae s.s. The performance of this new TaqMan assay was compared against the existing TaqMan assay and the standard PCR method in a blind species identification trial of over 450 samples using field collected specimens from a total of 13 countries in Sub-Saharan Africa. The standard PCR method was found to be specific with a low number of incorrect scores (<1%), however when compared to the TaqMan assays it showed a significantly higher number of failed reactions (15%). Both the new vector-specific TaqMan assay and the exisiting TaqMan showed a very low number of incorrectly identified samples (0 and 0.54%) and failed reactions (1.25% and 2.96%). In tests of analytical sensitivity the new TaqMan assay showed a very low detection threshold and can consequently be used on a single leg from a fresh or silica-dried mosquito without the need to first extract DNA. This study describes a rapid and sensitive assay that very effectively identifies the two main malaria vectors of the An. gambiae species complex from the non-vector sibling species. The method is based on TaqMan SNP genotyping and can be used to screen single legs from dried specimens. In regions where An. merus/melas/bwambae, vectors with restricted distributions, are not present it can be used alone to discriminate vector from non-vector or in combination with the Walker TaqMan assay to distinguish An. arabiensis and An. gambiae s.s.
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影响因子：
1.9
作者：
Fettene, M;Koekemoer, LL;Coetzee, M
通讯作者：
Coetzee, M
DOI：
10.1016/s0035-9203(98)90761-1
发表时间：
1998-03-01
影响因子：
2.2
作者：
Hunt, RH;Coetzee, M;Fettene, M
通讯作者：
Fettene, M
影响因子：
1.9
作者：
MAHON, RJ;GREEN, CA;HUNT, RH
通讯作者：
HUNT, RH
影响因子：
3
作者：
Wilkins, Elien E.;Howell, Paul I.;Benedict, Mark Q.
通讯作者：
Benedict, Mark Q.
影响因子：
3
作者：
Walker, Edward D.;Thibault, Alisha R.;Vulule, John M.
通讯作者：
Vulule, John M.