Aedes albopictus and aedes aegypti CYTB gene identification kit based on high-resolution melting curve and identification method thereof

Abstract

The invention relates to the field of species identification, and provides an aedes albopictus and aedes aegypti CYTB gene identification kit based on a high-resolution melting curve and an identification method thereof. The aedes albopictus and aedes aegypti CYTB gene identification kit based on the high-resolution melting curve is characterized in that universal primers are designed according toaedes albopictus and aedes aegypti CYTB gene sequences, and an optimized high-resolution melting curve reaction system is adopted to identify aedes albopictus and aedes aegypti of various forms suchas imagoes, ova, larvae and pupae. By the adoption of the aedes albopictus and aedes aegypti CYTB gene identification kit based on the high-resolution melting curve and the identification method thereof, aedes albopictus and aedes aegypti can be identified; operation is easy, the speed is high, no PCR products need post-treatment, the difference of a single base can be detected, and closed-tube operation is indeed achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a high-resolution melting curve-based identification kit for Aedes albopictus and Aedes aegypti. 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 populations such as closely related...

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

Images