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Loop-mediated isothermal amplification (LAMP) assays for Anopheles funestus group and Anopheles gambiae complex species.
Medical and Veterinary Entomology ( IF 1.6 ) Pub Date : 2020-03-10 , DOI: 10.1111/mve.12437 T Kamber 1 , L L Koekemoer 2, 3 , A Mathis 1
Medical and Veterinary Entomology ( IF 1.6 ) Pub Date : 2020-03-10 , DOI: 10.1111/mve.12437 T Kamber 1 , L L Koekemoer 2, 3 , A Mathis 1
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Species of the genus Anopheles vary with regard to their vector capacity for Plasmodium spp., the causative agent of malaria, and their accurate identification is often required. Loop‐mediated isothermal amplification (LAMP) is a rapid, simple and low‐cost method for specific DNA amplification. Primers for LAMP assays specific for the Anopheles funestus group and Anopheles gambiae complex species as well as for the species Anopheles arabiensis , An. funestus , An. gambiae s.s /Anopheles coluzzii (major vectors) and Anopheles rivulorum (minor vector) were designed targeting specific genome or rDNA internal transcribed spacer regions. Reaction conditions (buffer composition, primer concentrations, incubation time) were evaluated and the specificities of the assays confirmed with DNA from non‐target Anopheles species. DNA release from the mosquitoes is achieved simply by heating them for 5 min in water. An aliquot of the DNA solutions is transferred to the reaction tube using disposable inoculation loops. The outcome of the LAMP amplifications after 1 h incubation at 65 °C can easily be visualized by a colour change visible to the naked eye. The assays are operable under field conditions requiring only basic equipment (portable heat block programmable at 65 and 80 °C, cooler for master mixes).
中文翻译：
环状介导的等温扩增（LAMP）分析法用于按蚊群和冈比亚按蚊复杂物种。
按蚊属的种类根据其对疟疾的致病性疟原虫的载体能力而有所不同，通常需要对其进行准确鉴定。环介导的等温扩增（LAMP）是一种快速，简单且低成本的特异性DNA扩增方法。LAMP检测的引物，特定于真菌按蚊群和冈比亚按蚊复杂种以及阿拉伯按蚊（An按蚊）物种。funestus，一种。冈比亚ss /按蚊（主要媒介）和小按蚊（次要载体）被设计成靶向特定基因组或rDNA内部转录的间隔区。评估了反应条件（缓冲液组成，引物浓度，孵育时间），并用非靶标按蚊属物种的DNA确认了测定的特异性。从蚊子中释放DNA只需在水中加热5分钟即可。使用一次性接种环将等分试样的DNA溶液转移到反应管中。通过在肉眼可见的颜色变化可以很容易地看到在65°C下孵育1 h后LAMP扩增的结果。该测定法可在仅需基本设备的现场条件下进行操作（便携式加热块可在65和80°C进行编程，适用于预混液的冷却器）。
更新日期：2020-03-10
中文翻译：
环状介导的等温扩增（LAMP）分析法用于按蚊群和冈比亚按蚊复杂物种。
按蚊属的种类根据其对疟疾的致病性疟原虫的载体能力而有所不同，通常需要对其进行准确鉴定。环介导的等温扩增（LAMP）是一种快速，简单且低成本的特异性DNA扩增方法。LAMP检测的引物，特定于真菌按蚊群和冈比亚按蚊复杂种以及阿拉伯按蚊（An按蚊）物种。funestus，一种。冈比亚ss /按蚊（主要媒介）和小按蚊（次要载体）被设计成靶向特定基因组或rDNA内部转录的间隔区。评估了反应条件（缓冲液组成，引物浓度，孵育时间），并用非靶标按蚊属物种的DNA确认了测定的特异性。从蚊子中释放DNA只需在水中加热5分钟即可。使用一次性接种环将等分试样的DNA溶液转移到反应管中。通过在肉眼可见的颜色变化可以很容易地看到在65°C下孵育1 h后LAMP扩增的结果。该测定法可在仅需基本设备的现场条件下进行操作（便携式加热块可在65和80°C进行编程，适用于预混液的冷却器）。
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