{"title":"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","authors":"Srishty Raman , Pranay Tanwar , Jyoti Meena , Neerja Bhatla , Subhash C. Yadav","doi":"10.1016/j.sbsr.2024.100693","DOIUrl":null,"url":null,"abstract":"<div><p>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.</p></div>","PeriodicalId":424,"journal":{"name":"Sensing and Bio-Sensing Research","volume":"46 ","pages":"Article 100693"},"PeriodicalIF":5.4000,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214180424000758/pdfft?md5=74149227605c0a3b27f9a96f66efbae5&pid=1-s2.0-S2214180424000758-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Magnetic nanoparticles and quantum dots coupled immuno nano fluorescence assay for visual detection of HPV16-induced cervical cancer cells from cytology/biopsy samples\",\"authors\":\"Srishty Raman , Pranay Tanwar , Jyoti Meena , Neerja Bhatla , Subhash C. Yadav\",\"doi\":\"10.1016/j.sbsr.2024.100693\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>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.</p></div>\",\"PeriodicalId\":424,\"journal\":{\"name\":\"Sensing and Bio-Sensing Research\",\"volume\":\"46 \",\"pages\":\"Article 100693\"},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2024-09-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2214180424000758/pdfft?md5=74149227605c0a3b27f9a96f66efbae5&pid=1-s2.0-S2214180424000758-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Sensing and Bio-Sensing Research\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2214180424000758\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Sensing and Bio-Sensing Research","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2214180424000758","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
Magnetic nanoparticles and quantum dots coupled immuno nano fluorescence assay for visual detection of HPV16-induced cervical cancer cells from cytology/biopsy samples
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.
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.
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