快速鉴定四大家鱼早期资源种类的PCR方法

Species identification of early life of four major Chinese carps by PCR method

根据线粒体控制区序列设计了青鱼(Mylopharyngodon piceus)、草鱼(Ctenopharyngodon idellus)、鲢(Hypophthalmichthys molitrix)、鳙(Aristichthys nobilis)物种特异的PCR引物,通过对四大家鱼受精卵的PCR扩增,种间交叉扩增,相关种类早期资源的扩增以及未知种类鱼卵的鉴定发现,所设计的引物具有很高的种类特异性,对特定种DNA具有良好的扩增效果,但是不能进行交叉扩增,对四大家鱼外的其它种类也不能扩增。实验没有出现一例假阳性或假阴性结果,说明所设计引物的种类特异性高。该方法特别适合从鱼类早期资源大量样本中准确快速鉴定四大家鱼种类。

In China,Mylopharyngodon piceus,Ctenopharyngodon idellus,Hypophthalmichthys molitrix and Aristichthys nobilis are regarded as＂four major Chinese carps＂. In order to identify species of these carps quickly from large numbers of early life resource of river,we designed four species-specific PCR primers for the carps based on sequences of mitochondrial DNA control region. Validity of the primers was tested through samples in four major Chinese carps themselves,across them and some other common fish species from Yangtze River using PCR technology. The results of PCR showed that the aimed DNA bands were appeared clearly in primer-specific carps respectively but not amplified across four major Chinese carps,or in other common fish species. We also have taken positive and negative control in all PCR experiment,and no false results were found,which indicate that the designed species-primers worked effectively for four major Chinese carps.These primer sets is particularly suitable for rapidly identifying four major Chinese carp species from early life resources of ambiguous morphological characteristics.