Magnetic nanoparticles and quantum dots coupled immuno nano fluorescence assay for visual detection of HPV16-induced cervical cancer cells from cytology/biopsy samples

IF 5.4 Q1 CHEMISTRY, ANALYTICAL

Sensing and Bio-Sensing Research Pub Date : 2024-09-17 DOI:10.1016/j.sbsr.2024.100693

Srishty Raman , Pranay Tanwar , Jyoti Meena , Neerja Bhatla , Subhash C. Yadav

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引用次数: 0

Abstract

Biopsy-based histopathology and immunohistochemistry for cervical cancer detection are costly, time-consuming, and require expert personnel for data interpretation. We developed a simple magnetic nanoparticle (MNPs) and quantum dots (QD) coupled immuno nano fluorescence assay (MNPQDCINFA) for visual detection of HPV16-induced cervical cancer cells under UV light from cytology/biopsy samples exploiting host cancer cells expressing viral E7 protein as a biomarker. The E7 domain-specific polyclonal antibodies were generated against the 1–44 amino acid N-terminal (anti-domainN antibody) and 48–98 amino acid C-terminal domain (anti-domainC antibody). These antibodies were bioconjugated with nonfluorescent MNPs (60 % efficiency) and fluorescent QDs (66 % efficiency) to generate capturing (MNPs-anti-domainN antibody) and detecting (QDs-anti-domainC antibody) nano-complex, respectively. Assay conditions, such as concentration of capturing (20 μM) and detecting (50 nM) antibody nano-complexes and incubation duration (30 min), were standardized. The analytical sensitivity using pure HPV16 E7 protein was recorded up to 200 ng with very high specificity to differentiate from other HPV strains E7 proteins. The diagnostic performance characteristics with cytology samples showed 100 % sensitivity and specificity compared to immunofluorescence and biopsy-based histopathology analysis. The present invention can be effectively used for a quick, disposable, rapid cervical cancer cell detection system as an alternate test for immunofluorescence and histopathology.

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磁性纳米粒子和量子点耦合免疫纳米荧光检测法，用于从细胞学/活检样本中直观检测 HPV16 诱导的宫颈癌细胞

用于宫颈癌检测的活检组织病理学和免疫组织化学方法成本高、耗时长，而且需要专业人员进行数据解读。我们开发了一种简单的磁性纳米粒子（MNPs）和量子点（QD）耦合免疫纳米荧光检测法（MNPQDCINFA），利用表达病毒 E7 蛋白的宿主癌细胞作为生物标记物，在紫外光下从细胞学/活检样本中直观检测 HPV16 诱导的宫颈癌细胞。E7 结构域特异性多克隆抗体是针对 1-44 个氨基酸 N 端（抗结构域 N 抗体）和 48-98 个氨基酸 C 端（抗结构域 C 抗体）产生的。这些抗体分别与非荧光 MNPs（效率为 60%）和荧光 QDs（效率为 66%）进行生物共轭，生成捕获（MNPs-抗 domainN 抗体）和检测（QDs-抗 domainC 抗体）纳米复合物。捕获（20 μM）和检测（50 nM）抗体纳米复合物的浓度和孵育时间（30 分钟）等检测条件均已标准化。纯 HPV16 E7 蛋白的分析灵敏度高达 200 毫微克，特异性极高，可与其他 HPV 株 E7 蛋白区分开来。与免疫荧光和基于活检的组织病理学分析相比，细胞学样本的诊断性能特征显示出 100 % 的灵敏度和特异性。本发明可有效地用于快速、一次性、快速宫颈癌细胞检测系统，作为免疫荧光和组织病理学的替代检测方法。

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来源期刊

Sensing and Bio-Sensing Research Engineering-Electrical and Electronic Engineering

CiteScore

10.70

自引率

3.80%

发文量

审稿时长

87 days

期刊介绍： Sensing and Bio-Sensing Research is an open access journal dedicated to the research, design, development, and application of bio-sensing and sensing technologies. The editors will accept research papers, reviews, field trials, and validation studies that are of significant relevance. These submissions should describe new concepts, enhance understanding of the field, or offer insights into the practical application, manufacturing, and commercialization of bio-sensing and sensing technologies. The journal covers a wide range of topics, including sensing principles and mechanisms, new materials development for transducers and recognition components, fabrication technology, and various types of sensors such as optical, electrochemical, mass-sensitive, gas, biosensors, and more. It also includes environmental, process control, and biomedical applications, signal processing, chemometrics, optoelectronic, mechanical, thermal, and magnetic sensors, as well as interface electronics. Additionally, it covers sensor systems and applications, µTAS (Micro Total Analysis Systems), development of solid-state devices for transducing physical signals, and analytical devices incorporating biological materials.