Kit and method for identifying ND5 genes of culex pipiens pallens and culex fatigans based on high-resolution melting curve

Abstract

The invention relates to the field of species identification, and discloses a kit for identifying culex pipiens pallens and culex fatigans based on a high-resolution melting curve. The kit is characterized in that universal primers are designed according to the sequences of ND5 genes of the culex pipiens pallens and culex fatigans, and various forms such as bugs, eggs, larvae and pupae of the culex pipiens pallens and culex fatigans are identified through an optimized high-resolution melting curve reaction system. The culex pipiens pallens and culex fatigans can be identified, operation is simple, speed is high, no postprocessing is required by PCR products, the difference of a single basic group can be detected, and closed-tube operation is truly realized.

Description

technical field [0001] The invention relates to a kit for identifying Culex pipiens and Culex pipiens fatigued based on a high-resolution melting curve and an identification method thereof. It belongs to the technical field of biomedicine. Background technique [0002] The main means of identifying mosquito populations at ports is morphological identification, relying on port inspection and quarantine personnel who have experience in morphological identification and the ability to identify mosquito forms, and conduct identification according to the medical mosquito classification retrieval table. However, morphological identification faces many problems: there are few professional mosquito identification personnel, and the personnel training period is long; factors such as the experience level, professional level, mosquito integrity and developmental stage of identification personnel can lead to long identification period and low efficiency; It is difficult to identify popu...

AI Technical Summary

Problems solved by technology

However, morphological identification faces many problems: there are few professional mosquito identification personnel, and the personnel training period is long; factors such as the experience level, professional level, mosquito integrity and developmental stage of identification personnel can lead to long identification period and low efficiency; It is difficult to identify populations such as closely related populations and isomorphic populations; for mosquito eggs, larvae and pupae, in order to obtain their reliable identification characteristics, it is often necessary to identify mosquitoes after artificial cultivation; Class identification work is a very big test; etc.

However, these identification methods have shortcomings: routine detection of specific PCR product sequences relies on expensive molecular probes or product sequencing; RFLP identification of mosquito species is not only time-consuming, but also requires a large number of samples; RAPD identification of mosquito species is fast, but The results are unstable and sometimes irreproducible; SSCP can only be used as a method to detect mosquito gene mutations. To determine the location and type of mutations, further sequencing is required, and the electrophoresis conditions are relatively strict; the subsequent sequencing of DNA barcoding technology is time-consuming and labor-intensive. costly

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