A Rapid Identification Method of Bactocera cilifera (Hendel) with Species-Specific Primers (SS-COI)

[...]breeding of larvae and pupae intercepted is often limited by feeding time and different feeding conditions of fruit flies, which not only brings difficulties to rapid identification at port, but also affects fast customs clearance of import and export fruits and vegetables. [...]rapid identification method of different states of fruit flies is a technical problem urgently to be solved. [...]RFLP technique is complicated in operation, and identifies the interspecific relationship and species specificity of insects by detecting the variation of endonuclease recognition sites, with low polymorphism detection rate; RAPD technique has poor stability and is sensitive to the change of experimental environmental factors, and the amplified products have low repeatability and stability, which limit the wide application of RFLP and RAPD technique[lcl Although AFLP technology better makes up for the deficiencies of the above two methods, it has high requirements on DNA quality of detection samples, and the cost of required equipment is relatively expensive. [...]the above detection methods are difficult to meet the requirements of fast customs clearance, accurate identification and low-cost quarantine for import and export trade of fruits and vegetables at ports. Because mtDNA COI gene is relatively conservative, with relatively stable and close arrangement, it is easily amplified by universal primers, and has enough specificity to distinguish different species. According to the sequences of B" cilifera (Hendel), the sequences with high homology were screened by BLAST program in NCBI database.