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目的建立一种快速、灵敏、特异的用于检测贝类奥尔森派琴虫的实时荧光定量PCR方法。方法根据基因库中奥尔森派琴虫的基因保守序列,设计合成1对引物和1条TaqMan探针,建立荧光定量PCR方法,对采自广西沿海的49份贻贝标本进行检测,并与常规PCR比较。结果建立的荧光定量PCR方法灵敏度可达20个拷贝,比常规PCR灵敏度高100倍。49份贻贝标本的阳性率为16.3%,检测的奥尔森派琴虫基因组DNA含量为2.38×106～9.21×102拷贝/μl。结论建立的荧光定量PCR方法可以用于贝类奥尔森派琴虫感染的快速检测。
Objective To establish a rapid, sensitive and specific real-time PCR method for detection of shellfish Olson flies. Methods According to the conserved sequence of Olsen fusorum in the gene bank, one pair of primers and one TaqMan probe were designed and synthesized, and 49 samples of mussels collected from the coast of Guangxi were detected by fluorescence quantitative PCR. Conventional PCR comparison. Results The fluorescence quantitative PCR method was established with a sensitivity of 20 copies, which is 100 times more sensitive than conventional PCR. The positive rate of 49 mussel specimens was 16.3%. The genomic DNA content of Olszpf isolated worm was 2.38 × 106 ~ 9.21 × 102 copies / μl. Conclusion The established fluorescence quantitative PCR method can be used for the rapid detection of shellfish Olson fidbits.