奥尔森派琴虫PCR检测方法的优化及其在进出境检疫中的应用

Optimization of PCR detection for Perkinsus olseni and its application in entry and exit quarantine

为快速、准确地检测进出境水产品中的派琴虫,防止有害寄生虫传入,对OIE推荐的奥尔森派琴虫PCR检测方法的扩增程序进行优化,并应用于上海口岸从16个国家进口的21种贝类水产品95批次的奥尔森派琴虫的检疫。结果显示,该优化的PCR扩增程序用时由原来的224 min缩短为91 min,效率提高了2.6倍,且最低检测限相同;并从澳大利亚进口的9个扇贝样品中检出7份奥尔森派琴虫阳性,阳性率为77.8%。测序结果表明,与奥尔森派琴虫序列(登录号为U07701.1)的同源性为99%。试验证明优化的PCR检测方法快速、准确,适用于进出境水产品中奥尔森派琴虫的检疫,并提示在进口贝类水产品中,应加强澳大利亚进口水产品的检疫。

To establish a rapid and specific PCR method to detect Perkinsus olseni and prevent this harmful parasite from spreading into the country,the PCR detection method recommended by the World 0rganisation forinimal Health（OIE） was optimized and then applied for detection of 95 batches of samples collected from 21 types of shellfish products from 16 countries. The result showed that without the change of sensibility, the optimized PCR procedure has decreased the time from 224 min to 91 man. The efficiency of the optimized PCR procedure has increased to 2.6 times. ~ olseni was detected in 77.8% （7/9） scallops imported from Australia. The sequence analysis revealed that the homology was 99% （U07701.1）.The results showed that the optimized PCR method met requirements for use in rapid testing of P.olseni. Moreover the aquatic products imported from Australia should be detected carefully.