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Dual platform based sandwich assay surface-enhanced Raman scattering DNA biosensor for the sensitive detection of food adulteration.
Analyst ( IF 3.6 ) Pub Date : 2019-12-17 , DOI: 10.1039/c9an02106j
Ibrahim Khalil 1 , Wageeh A Yehye , Nurhidayatullaili Muhd Julkapli , Abu Ali Ibn Sina , Shahrooz Rahmati , Wan Jefrey Basirun , Ali Seyfoddin
Affiliation  

Surface enhanced Raman scattering (SERS) DNA biosensing is an ultrasensitive, selective, and rapid detection technique with the ability to produce molecule-specific distinct fingerprint spectra. It supersedes the long amplicon based PCR assays, the fluorescence and spectroscopic techniques with their quenching and narrow spectral bandwidth, and the electrochemical detection techniques using multiplexing. However, the performance of the SERS DNA biosensor relies on the DNA probe length, platform composition, both the presence and position of Raman tags and the chosen sensing strategy. In this context, we herein report a SERS biosensor based on dual nanoplatforms with a uniquely designed Raman tag (ATTO Rho6G) intercalated short-length DNA probe for the sensitive detection of the pig species Sus scrofa. In the design of the signal probe (SP), a Raman tag was incorporated adjacent to the spacer arm, followed by a terminal thiol modifier, which consequently had a strong influence on the SERS signal enhancement. The detection strategy involves the probe-target DNA hybridization mediated coupling of the two platforms, i.e., the graphene oxide-gold nanorod (GO-AuNR) functionalized capture probe (CP) and SP-conjugated gold nanoparticles (AuNPs), consequently enhancing the SERS intensity by both the electromagnetic hot spots generated at the junctions or interstices of the two platforms and the chemical enhancement between the AuNPs and the adsorbed intercalated Raman tag. This dual platform based SERS DNA biosensor exhibited outstanding sensitivity in detecting pork DNA with a limit of detection (LOD) of 100 aM validated with DNA extracted from a pork sample (LOD 1 fM). Moreover, the fabricated SERS biosensor showed outstanding selectivity and specificity for differentiating the DNA sequences of six closely related non-target species from the target DNA sequences with single and three nucleotide base-mismatches. Therefore, the developed short-length DNA linked dual platform based SERS biosensor could replace the less sensitive traditional methods of pork DNA detection and be adopted as a universal detection approach for the qualitative and quantitative detection of DNA from any source.

中文翻译:

基于双平台的三明治检测表面增强拉曼散射DNA生物传感器,用于食品掺假的灵敏检测。

表面增强拉曼散射(SERS)DNA生物传感是一种超灵敏,选择性和快速的检测技术,具有产生特定于分子的独特指纹光谱的能力。它取代了基于长扩增子的PCR测定法,具有猝灭和窄光谱带宽的荧光和光谱技术以及使用多路复用的电化学检测技术。但是,SERS DNA生物传感器的性能取决于DNA探针的长度,平台组成,拉曼标签的存在和位置以及所选择的传感策略。在此背景下,我们在此报告了一种基于双纳米平台的SERS生物传感器,该传感器具有独特设计的拉曼标签(ATTO Rho6G)插入的短长度DNA探针,用于灵敏地检测猪种Sus scrofa。在信号探头(SP)的设计中,在间隔臂附近掺入拉曼标签,然后加入末端硫醇改性剂,因此对SERS信号增强具有很强的影响。该检测策略涉及探针-靶标DNA杂交介导的两个平台的偶联,即氧化石墨烯-金纳米棒(GO-AuNR)功能化捕获探针(CP)和SP偶联金纳米颗粒(AuNPs),从而增强了SERS在两个平台的交界处或空隙处产生的电磁热点,以及AuNP与吸附的拉曼标签之间的化学增强作用,都可产生较高的强度。这种基于双平台的SERS DNA生物传感器在检测猪肉DNA方面表现出出色的灵敏度,检测限(LOD)为100 aM,并用从猪肉样品中提取的DNA(LOD 1 fM)进行了验证。而且,所制备的SERS生物传感器显示出出色的选择性和特异性,可将6个密切相关的非靶标物种的DNA序列与具有单核苷酸和3个核苷酸碱基错配的目标DNA序列区分开。因此,开发的基于短链DNA链接的双平台SERS生物传感器可以代替敏感性较低的传统猪肉DNA检测方法,并可以作为一种通用的检测方法,用于对任何来源的DNA进行定性和定量检测。
更新日期:2020-02-17
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