Microfluid biosensor for detection of hpv in patient scraping samples: Determining E6 and E7 oncogenes

IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Bioelectrochemistry Pub Date : 2024-08-13 DOI:10.1016/j.bioelechem.2024.108795
Léony S. Oliveira , Sanket Goel , Khairunnisa Amreen , Norma Lucena-Silva , Maria D.L. Oliveira , Cesar A.S. Andrade
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Abstract

E6 and E7 oncogenes are pivotal in the carcinogenic transformation in HPV infections and efficient diagnostic methods can ensure the detection and differentiation of HPV genotype. This study describes the development and validation of an electrochemical, label-free genosensor coupled with a microfluidic system for detecting the E6 and E7 oncogenes in cervical scraping samples. The nanostructuring employed was based on a cysteine and graphene quantum dots layer that provides functional groups, surface area, and interesting electrochemical properties. Biorecognition tests with cervical scraping samples showed differentiation in the voltammetric response. Low-risk HPV exhibited a lower biorecognition response, reflected in ΔI% values of 82.33 % ± 0.29 for HPV06 and 80.65 % ± 0.68 for HPV11 at a dilution of 1:100. Meanwhile, high-risk, HPV16 and HPV18, demonstrated ΔI% values of 96.65 % ± 1.27 and 93 % ± 0.026, respectively, at the same dilution. Therefore, the biorecognition intensity followed the order: HPV16 >HPV18 >HPV06 >HPV11. The limit of detection and the limit of quantification of E6E7 microfluidic LOC−Genosensor was 26 fM, and 79.6 fM. Consequently, the E6E7 biosensor is a valuable alternative for clinical HPV diagnosis, capable of detecting the potential for oncogenic progression even in the early stages of infection.

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用于检测患者刮片样本中 hpv 的微流控生物传感器:确定 E6 和 E7 致癌基因
E6和E7致癌基因在人乳头瘤病毒感染的癌变过程中起着关键作用,高效的诊断方法可以确保检测和区分人乳头瘤病毒的基因型。本研究介绍了一种电化学无标记基因传感器的开发和验证,该传感器与微流控系统相结合,用于检测宫颈刮片样本中的 E6 和 E7 致癌基因。采用的纳米结构基于半胱氨酸和石墨烯量子点层,可提供功能基团、表面积和有趣的电化学特性。宫颈刮片样本的生物识别测试表明,伏安反应存在差异。低危 HPV 的生物识别响应较低,在 1:100 稀释度下,HPV06 和 HPV11 的 ΔI% 值分别为 82.33 % ± 0.29 和 80.65 % ± 0.68。与此同时,在相同的稀释度下,高危型 HPV16 和 HPV18 的 ΔI% 值分别为 96.65 % ± 1.27 和 93 % ± 0.026。因此,生物识别强度依次为HPV16>HPV18>HPV06>HPV11。E6E7 微流控 LOC-Genosensor 的检测限和定量限分别为 26 fM 和 79.6 fM。因此,E6E7 生物传感器是临床人类乳头瘤病毒诊断的一种有价值的替代方法,即使在感染的早期阶段也能检测到潜在的致癌进展。
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来源期刊
Bioelectrochemistry
Bioelectrochemistry 生物-电化学
CiteScore
9.10
自引率
6.00%
发文量
238
审稿时长
38 days
期刊介绍: An International Journal Devoted to Electrochemical Aspects of Biology and Biological Aspects of Electrochemistry Bioelectrochemistry is an international journal devoted to electrochemical principles in biology and biological aspects of electrochemistry. It publishes experimental and theoretical papers dealing with the electrochemical aspects of: • Electrified interfaces (electric double layers, adsorption, electron transfer, protein electrochemistry, basic principles of biosensors, biosensor interfaces and bio-nanosensor design and construction. • Electric and magnetic field effects (field-dependent processes, field interactions with molecules, intramolecular field effects, sensory systems for electric and magnetic fields, molecular and cellular mechanisms) • Bioenergetics and signal transduction (energy conversion, photosynthetic and visual membranes) • Biomembranes and model membranes (thermodynamics and mechanics, membrane transport, electroporation, fusion and insertion) • Electrochemical applications in medicine and biotechnology (drug delivery and gene transfer to cells and tissues, iontophoresis, skin electroporation, injury and repair). • Organization and use of arrays in-vitro and in-vivo, including as part of feedback control. • Electrochemical interrogation of biofilms as generated by microorganisms and tissue reaction associated with medical implants.
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