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根据锦鲤疱疹病毒(koi herpesvirus,KHV)的基因序列合成了2对引物,对送检的发病锦鲤样品提取DNA进行PCR检测,分别扩增出KHV的胸腺嘧啶脱氧核苷激酶基因(thymidine kinase,TK)409 bp和AF411803基因序列484 bp的条带,对484 bp的PCR扩增产物经限制性内切酶TaqⅠ和AluⅠ酶切,分别得到200 bp/284 bp和185 bp/299 bp的片段,484 bp的PCR扩增产物的测序结果与GenBank上已发表的KHV序列一致。结果显示发病锦鲤为KHV阳性。
Two pairs of primers were synthesized according to the gene sequence of koi herpesvirus (KHV). PCR was carried out on the DNA extracted from the diseased Koi samples. The thymidine kinase gene of KHV (thymidine kinase) , TK) 409 bp and AF411803 484 bp, respectively. The 484 bp PCR products were digested with restriction endonucleases Taq Ⅰ and Alu Ⅰ to obtain fragments of 200 bp / 284 bp and 185 bp / 299 bp, respectively The sequencing result of the 484 bp PCR amplification product was consistent with the published KHV sequence in GenBank. The results showed that Koi positive for KHV.