Abstract:
The current study was conducted to statistically compare the SYBR® Green quantitative polymerase chain reaction (qPCR) assay and the conventional plate counting (PC) method to construct growth curves of a cocktail of Weissella viridescens in pure culture under different isothermal storage conditions (4, 8, 14, and 30 °C) and in mixed culture with Leuconostoc mesenteroides at 8 °C. The efficiency and specificity of the qPCR standard curves were confirmed, and both methods were adequate to quantify the growth kinetics of W. viridescens at all isothermal temperatures, demonstrating a good correlation and agreement. The efficiencies of the standard curves varied between 98% and 102%. The SYBR® Green qPCR assay was also able to differentiate the growth curves of W. viridescens and L. mesenteroides in the mixed culture at 8 °C. Additionally, the SYBR® Green qPCR method was considered a faster and more sensitive alternative to construct growth curves under different isothermal conditions and differentiate morphologically similar lactic acid bacteria. Overall, the results suggest that the SYBR® Green qPCR method is a reliable and efficient tool to study microbial growth kinetics in pure and mixed cultures.
摘要:
进行本研究以统计学比较SYBR®Green定量聚合酶链反应(qPCR)测定法和常规平板计数(PC)方法,以构建在不同等温储存条件(4、8、14和30°C)下的纯培养物中的Weissella病毒基因混合物的生长曲线,并在8°C下与串珠菌混合培养qPCR标准曲线的效率和特异性得到证实,这两种方法都足以量化在所有等温温度下紫草的生长动力学,表现出良好的相关性和一致性。标准曲线的效率在98%和102%之间变化。SYBR®GreenqPCR测定还能够区分8°C下混合培养物中病毒基因W.viridesecens和中肠曲霉菌的生长曲线。此外,SYBR®GreenqPCR方法被认为是在不同等温条件下构建生长曲线并区分形态相似的乳酸菌的更快,更灵敏的替代方法。总的来说,结果表明,SYBR®GreenqPCR方法是研究纯培养物和混合培养物中微生物生长动力学的可靠且有效的工具。