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Abstract：
Bacterial pathogens pose an increasing food safety and bioterrorism concern. Current DNA detection methods utilizing sensitive nanotechnology and biosensors have shown excellent detection, but require expensive and time-consuming polymerase chain reaction (PCR) to amplify DNA targets; thus, a faster, more economical method is still essential. In this proof-of-concept study, we investigated the ability of a gold nanoparticle-DNA (AuNP-DNA) biosensor to detect non-PCR amplified genomic Salmonella enterica serovar Enteritidis (S. enteritidis) DNA, from pure or mixed bacterial culture and spiked liquid matrices. Non-PCR amplified DNA was hybridized into sandwich-like structures (magnetic nanoparticles/DNA/AuNPs) and analyzed through detection of gold voltammetric peaks using differential pulse voltammetry. Our preliminary data indicate that non-PCR amplified genomic DNA can be detected at a concentration as low as 100 ng/mL from bacterial cultures and spiked liquid matrices, similar to reported PCR amplified detection levels. These findings also suggest that AuNP-DNA biosensors are a first step towards a viable detection method of bacterial pathogens, in particular, for resource-limited settings, such as field-based or economically limited conditions. Future efforts will focus on further optimization of the DNA extraction method and AuNP-biosensors, to increase sensitivity at lower DNA target concentrations from food matrices comparable to PCR amplified DNA detection strategies.
摘要：
细菌病原体引起越来越多的食品安全和生物恐怖主义问题。目前利用灵敏的纳米技术和生物传感器的DNA检测方法已显示出优异的检测性能,但需要昂贵且耗时的聚合酶链反应（PCR）来扩增DNA靶标；因此，更快,更经济的方法仍然是必不可少的。在这个概念验证研究中,我们研究了金纳米颗粒-DNA（AuNP-DNA）生物传感器检测非PCR扩增的基因组肠道沙门氏菌的能力（S.肠炎)DNA，来自纯或混合细菌培养物和加标液体基质。将非PCR扩增的DNA杂交成夹心样结构(磁性纳米颗粒/DNA/AuNP),并通过使用差分脉冲伏安法检测金伏安峰来分析。我们的初步数据表明，可以从细菌培养物和加标液体基质中检测到浓度低至100ng/mL的非PCR扩增的基因组DNA，与报告的PCR扩增检测水平相似。这些发现还表明，AuNP-DNA生物传感器是细菌病原体可行检测方法的第一步，特别是,对于资源有限的设置，例如基于现场或经济有限的条件。未来的工作将集中在进一步优化DNA提取方法和AuNP-生物传感器，与PCR扩增的DNA检测策略相比，可以在较低的食物基质DNA靶浓度下提高灵敏度。
