一种基于纳米颗粒的轮状病毒免疫检测方法的开发和评估:在低收入环境中替代ELISA和PCR的合适方法。

IF 2 Q3 BIOCHEMICAL RESEARCH METHODS
Margaret Oluwatoyin Japhet, Adeogo Timilehin Bankole, Temiloluwa Ifeoluwa Omotade, Oyelola Eyinade Adeoye, Oladiran Famurewa, Simeon K Adesina
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引用次数: 0

摘要

每年,腹泻造成100万0至5岁儿童死亡,其中轮状病毒是主要原因。受影响最严重的地区由于费用高、缺乏必要的设备和缺乏酶联免疫吸附试验(ELISA)和分子方法的训练有素的人员,缺乏常规的轮状病毒诊断。我们报道了一种廉价的、基于纳米颗粒的常规轮状病毒诊断免疫测定方法的开发和评估。本文利用傅里叶变换红外光谱(FTIR)确定了棉签氧化和醛生产试剂盒的最佳条件。将将病毒与棉签结合所需的乳铁蛋白(LF)固定在活化的棉签上,然后在LF固定的棉签上捕获商品轮状病毒抗原。将其浸在与轮状病毒群特异性单克隆抗体共价偶联的有色纳米粒中,用于轮状病毒视觉检测。随后,对同一组186份粪便样本进行纳米法、商用ELISA和定量反转录PCR检测轮状病毒的比较,并进行统计分析。在0.1 M醋酸钠缓冲液中,用48 mg/mL的NaIO4在35°C下氧化9 h,观察到最佳氧化条件。多次洗涤后,通过棉签上的蓝色保留,肉眼证实了轮状病毒的检测。ELISA检测轮状病毒的灵敏度、特异性、阳性预测值和阴性预测值分别为60%、84%、53%和88%,而我们的免疫分析法检测轮状病毒的灵敏度、特异性、阳性预测值和阴性预测值分别为88%、94%、82%和96%。这种免疫分析将为发病率/死亡率高的中低收入国家提供有效的轮状病毒公共卫生干预措施。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Development and Evaluation of a Nanoparticle-Based Immunoassay for Rotavirus Detection: A Suitable Alternative to ELISA and PCR in Low-Income Setting.

Development and Evaluation of a Nanoparticle-Based Immunoassay for Rotavirus Detection: A Suitable Alternative to ELISA and PCR in Low-Income Setting.

Development and Evaluation of a Nanoparticle-Based Immunoassay for Rotavirus Detection: A Suitable Alternative to ELISA and PCR in Low-Income Setting.

Development and Evaluation of a Nanoparticle-Based Immunoassay for Rotavirus Detection: A Suitable Alternative to ELISA and PCR in Low-Income Setting.

Every year, diarrhoea is responsible for >1 million deaths in children with ages from 0 to 5 years, with rotavirus as the leading cause. The regions most affected lack routine rotavirus diagnosis due to high cost, lack of necessary equipment and shortage of trained-personnel for Enzyme-Link-Immunosorbent-Assay (ELISA) and molecular methods. We report the development and evaluation of a cheap, nanoparticle-based immunoassay for routine machine-free rotavirus diagnosis. In this work, optimal conditions for oxidation of cotton swabs and aldehyde production for kit development was confirmed by Fourier-Transform Infrared Spectroscopy (FTIR). Lactoferrin (LF) needed to bind the virus to the cotton swab was immobilised on activated cotton swabs, followed by the capture of commercial rotavirus antigen on LF-immobilised swabs. This was dipped in coloured nanobeads covalently coupled to rotavirus-group-specific monoclonal antibody for visual rotavirus detection. Subsequently, rotavirus detection by nanoassay, commercial ELISA and quantitative reverse transcription PCR were compared using same set of 186 stool samples and subjected to statistical analyses. Optimal oxidisation condition was observed using 48 mg/mL NaIO4 in 0.1 M sodium acetate buffer at 35 °C for 9 h. Rotavirus detection was confirmed visually by blue colour retention on swabs after several washings. Sensitivity, specificity, positive-predictive-value and negative-predictive-value of ELISA in rotavirus detection were 60%, 84%, 53% and 88%, respectively, while our immunoassay showed performance at 88%, 94%, 82% and 96%. This immunoassay will provide effective rotavirus public health interventions in low-and-middle-income countries with high morbidity/mortality.

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来源期刊
Methods and Protocols
Methods and Protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
3.60
自引率
0.00%
发文量
85
审稿时长
8 weeks
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