Allglo probe-based detection method of anopheles sinensis knockdown resistance gene mutation site

Abstract

The invention discloses an Allglo probe-based detection method of an anopheles sinensis knockdown resistance gene mutation site. A special primer provided by the invention consists of a primer 1 and a primer 2 shown in a SEQ ID No.1 and a SEQ ID No.2; a reagent consists of the special primer, a probe 1 (sequence SEQ ID No.3), a probe 2 (sequence SEQ ID No.4) and a probe 3 (sequence SEQ ID No.4); a reaction reagent consists of a reagent and an Allglo reaction buffer liquid; and a kit comprises the reaction reagent. The invention provides an efficient, rapid, simple and convenient anopheles sinensis kdr gene mutation site detecting method which is high in specificity and sensitivity, and according to the method, the anopheles sinensis kdr gene mutation type can be rapidly and accurately detected.

Description

technical field [0001] The invention belongs to the technical field of gene mutation detection, in particular to a detection method for anopheles sinensis knockdown resistance (kdr) gene mutation site, in particular to an Anopheles sinensis knockdown resistance (kdr) gene mutation site based on Allglo probe Fluorescent quantitative PCR detection method and kit. Background technique [0002] Anopheles sinensis is one of the main malaria vectors in my country. Recent reports indicate that the transmissibility of Anopheles sinensis in the central part of China has been greatly improved compared with the last century. Malaria outbreaks in central my country and the Huanghuai region are closely related. The use of insecticides to control Anopheles mosquitoes is a very important means of malaria prevention and control. Humans have used insecticides to control mosquitoes for more than 80 years. Since the early 1980s, pyrethroid insecticides have been used to treat mosquito nets and...

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Problems solved by technology

In 2011, Wu Song et al. reported the establishment of PCR detection method for knockdown resistance mutation of Anopheles sinensis (Wu Song, Ma Jian, Liu Qian et al. Establishment of PCR detection method for knockdown resistance mutation of Anopheles sinensis[J]. Journal of Comorbid Diseases, 2011, 27(11): 1008-1010); Chinese patents CN102373280A and CN102373282A respectively disclose a kind of Anopheles sinensis drug resistance AS-PCR detection kit and PCR-RFLP detection kit, but the above PCR method Need to open the cover to test the results, which is easy to cause cross-contamination and false positives. The result needs to be judged according to the strength of the electrophoresis band, which reduces the sensitivity and specificity of the test, and is easy to misjudgment and misjudgment. It is not conducive to the health of operators; Chinese patent CN102373281A discloses a TaqMan PCR detection kit for Anopheles sinensis drug resistance, but the synthesis of TaqMan probes is complicated and expensive, and it is necessary to use different TaqMan probe combinations in two tubes to perform fluorescence quantitative amplification respectively. The operation and detection procedures are cumbersome, and the detection sensitivity and specificity need to be improved; in addition, the above methods need to pre-extract Anopheles mosquito genome DNA as the amplification template, which is time-consuming, labor-intensive, low detection efficiency, and easy to contaminate

[0005] In the on-site vector monitoring, there is still a lack of high sensitivity and specificity, low cost, simple and fast detection method of kdr gene mutation of Anopheles sinensis and corresponding reagents and kits

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