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在食品中同时检测和定量的ddPCR方法

Xueqi Sun¹, Jian Xiao², Haiyan Wei³

¹Guangzhou Food Inspection Institute, Guangzhou, 511400, Guangdong, China. sun-xueqi@163.com.

Scientific reports

|September 1, 2025

概括

这项研究引入了快速四倍滴滴数字聚合酶链反应 (ddPCR) 方法,用于检测四种主要的食源细菌. 与传统方法相比,ddPCR测定具有更高的速度,灵敏度和可重复性.

科学领域:

食品安全问题
微生物学
分子诊断

背景情况:

传统的食物传播病原体检测速度缓慢且容易出现错误.
准确量化病原体对于预防食物传播疾病至关重要.
滴滴数字PCR (ddPCR) 提供了高精度的分子量化.

研究的目的:

开发一个四重的ddPCR试验,同时检测和定量四种主要的食源病原体.
评估 ddPCR 方法的准确性和灵敏性.
将ddPCR方法与传统培养技术进行比较.

主要方法:

开发针对沙门氏菌,黄金葡萄球菌,单细胞菌和白菌的多重ddPCR测定方法.
在定义的病原体度范围内使用ddPCR进行量化.
使用人工污染的食品样本进行验证,并与盘子计数进行比较.

主要成果:

在所有四种病原体的测试范围中,ddPCR测试显示出强烈的线性 (r2 > 0. 999).
检测的低极限是:8副本/20μL (S. Typhi),7副本/20μL (S. aureus),9副本/20μL (L. monocytogenes) 和7副本/20μL (B. cereus).
结果显示与盘子计数相比没有统计差异,但在速度,检测极限和可重复性方面有显著改善.

结论:

已建立的四重dPCR方法提供了一个快速,灵敏和可重复的工具来检测多种食源性病原体.
这种先进的分子技术在食品安全应用中的效率和准确性超越了传统方法.
ddPCR测定对常规查和控制食源性疾病具有重大潜力.

关键词:

类植物李斯特菌单细胞菌沙门氏菌黄金葡萄球菌通过食物传播的病原体四重点滴滴数字PCR (ddPCR)

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PCR - Polymerase Chain Reaction

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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