Development of a Microfluidics-Based Quantitative Real-Time PCR to Rapidly Identify Photobacterium damselae subsp. damselae with Different Pathogenicity by Detecting the Presence of mcp or dly Gene
文献类型： 外文期刊
作者： Zhang Zheng 1 ; Yu Yongxiang 1 ; Chen Jing 1 ; Wang Yingeng 1 ; Jiang Yong 3 ; Liao Meijie 1 ; Rong Xiaojun 1 ; Zhang Hao 1 ;
作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Key Lab Maricultural Organism Dis Control, Minist Agr & Rural Affairs, Qingdao 266071, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266237, Peoples R China
3.Natl Oceanog Ctr, Qingdao 266071, Peoples R China
关键词： mariculture; Photobacterium damselae; microfluidics; pathogenicity; rapid detection
期刊名称：JOURNAL OF OCEAN UNIVERSITY OF CHINA （ 影响因子：0.913； 五年影响因子：1.012 ）
ISSN： 1672-5182
年卷期： 2021 年 20 卷 2 期
页码：
收录情况： SCI
摘要： As a marine bacterial pathogen, Photobacterium damselae subsp. damselae (PDD) is distributed in seawater worldwide. It can infect different animals as well as humans, even cause deaths. The highly conserved regions of PDD mcp gene on chromosome and dly gene on plasmid were selected as the target fragments to design the specific primers. Recombinant plasmid standard was prepared based on the primers. With GENECHECKER UF-150 qRT-PCR instrument as the platform, a fluorescence-based quantitative real-time PCR (qRT-PCR) method was established for the detection of PDD. This method can specifically detect PDD and distinguish the highly virulent strains. Additionally, the test results can be qualitatively judged by visualization, while the quantitative detection can be achieved through the standard curve calculation. The minimum limit of detection was 1.0x10(1) copies mu L-1, and the detection time was less than 20 min. In summary, this new method has outstanding advantages, such as strong specificity, high sensitivity, and low site requirements. It is a rapid on-site detection technology for highly virulent PDD strains.
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