PCR方法快速鉴定两种检疫性豆象

采用形态学观察与PCR技术相结合的方法,针对口岸截获的可疑菜豆象和四纹豆象幼虫和成虫进行快速鉴定。采用异硫氰酸胍方法提取昆虫基因组DNA,并根据特异性引物,采用PCR技术从截获的可疑菜豆象幼虫和成虫中均扩增得到192 bp的特异性条带,从截获的可疑四纹豆象幼虫和成虫中均扩增得到181 bp的特异性条带,经测序比对,扩增产物序列与菜豆象和四纹豆象Cyt b目的基因序列完全相同。由此证实可疑样本即为菜豆象和四纹豆象的幼虫和成虫。该方法缩短了鉴定周期,6 h内即可快速准确鉴定未成熟的虫态,为口岸检验检疫及快速通关提供依据。

Rapid Identification of Two Quarantine Bean Weevil with PCR Method

Morphological observation combined with PCR methods were used for the rapid quarantine and identification of dubious larvae and adult of Acanthoscelides obtectus and Callosobruchus maculatus from port.From the genomic DNA of the two dubious samples,we amplified and sequenced a 192 bp and 181 bp PCR special product respectively,which had an identical sequence to the target gene Cyt b in Acanthoscelides obtectus and Callosobruchus maculatus respectively.These results proved that the above two dubious samples were the larvae and adult of Acanthoscelides obtectus and Callosobruchus maculatus.This method shortened the identification period,and could rapidly and accurately identify the immature insect-state of Acanthoscelides obtectus and Callosobruchus maculatus within six hours,so it should be useful for practical application in port of entry quarantine.