哈维氏弧菌
生物
DNA提取
溶藻弧菌
实时聚合酶链反应
聚合酶链反应
底漆(化妆品)
微生物学
弧菌科
检出限
基因
弧菌
色谱法
遗传学
细菌
化学
有机化学
作者
Cátia Costa,Guilherme Duarte Ferreira,Marco Simões,Joana Silva,Maria Jorge Campos
出处
期刊:Microorganisms
[MDPI AG]
日期:2022-10-18
卷期号:10 (10): 2060-2060
被引量:3
标识
DOI:10.3390/microorganisms10102060
摘要
Vibriosis, an often-fatal disease induced by pathogenic members of the Vibrionaceae family, causes severe economic losses in aquacultures. To mitigate/avoid vibriosis outbursts, it is vital to detect and quantify these pathogens as early as possible. However, standard microbiological methods are time-consuming and often underestimate cell counts, which calls for the development of valid alternatives. In this study, real-time polymerase chain reaction (qPCR) was employed to detect the pathogenic species Vibrio alginolyticus, Listonella anguillara, and Vibrio harveyi using a new primer pair targeting the groEL gene. In addition, the DNA extraction efficiency of three methods, two commercial kits and the boiling method, was compared. The most efficient method was the DNeasy Blood and Tissue kit, with a detection limit ranging between 154 and 600 CFU mL-1 in the case of V. alginolyticus and L. anguillara, and 48 CFU mL-1 for V. harveyi. Thus, this study presents the development and evaluation of a method for the early quantification of all three species in saline suspensions. However, the results obtained by spiking a microalgae sample with V. harveyi emphasize the importance of adjusting the DNA control's standard curve to the relevant extraction matrices, as it affects the DNA extraction efficiency and may hamper an accurate quantification with qPCR.
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