珊瑚病原菌株XSBZ03和XSBZ14双重PCR检测方法的建立

Duplex PCR assay for detection of coral pathogenic strains XSBZ03 and XSBZ14

摘要:

[目的]建立珊瑚病原菌株XSBZ03和XSBZ14的双重PCR检测方法。[方法]以XSBZ03和XSBZ14的特异靶序列为对象，开展引物设计和双重PCR检测方法的构建，并确定该双重PCR方法的特异性、敏感性及可靠性。[结果]该检测方法可特异识别菌株XSBZ03和XSBZ14，对XSBZ03和XSBZ14基因组DNA样品的检测极限分别为1.7 pg/μL和2.0 pg/μL；对XSBZ03和XSBZ14在海水样品中的检测极限分别为6×10³ CFU/mL和8×10³ CFU/mL。[结论]该方法具有特异性强、灵敏度高等优点，可对由菌株XSBZ03和XSBZ14引起的珊瑚疾病进行准确快速的诊断，为今后开展珊瑚疾病防控和无特定病原的珊瑚移植提供了可靠手段。

关键词:珊瑚病原;双重PCR;诊断

Abstract:

[Objective] To establish a duplex PCR assay for detection of the strains XSBZ03 and XSBZ14. [Methods] Strain-specific primers were designed according to the target sequence of XSBZ03 and XSBZ14. Subsequently, duplex PCR assay based on the primers was successfully established. [Results] The duplex PCR assay could detect accurately strains XSBZ03 and XSBZ14. The detection limit for XSBZ03 and XSBZ14 genomic DNA concentration was 1.7 pg/μL and 2.0 pg/μL. In artificial seawater samples, the detection limit for XSBZ03 and XSBZ14 strain concentration was 6×10³ CFU/mL and 8×10³ CFU/mL, respectively. [Conclusion] This assay provides a reliable method for the rapid diagnosis of Porites andrewsi White Syndrome (PAWS) and the Specific Pathogen Free (SPF) coral transplantation caused by these two pathogens.

Key words:coral pathogen;duplex PCR;diagnosis