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利用荧光胶束硅胶纳米传感器研制免疫层析条检测试剂盒,用于快速检测牛奶样品中的牛布鲁氏菌抗体。

Development of immunochromatographic strip test using fluorescent, micellar silica nanosensors for rapid detection of B. abortus antibodies in milk samples.

机构信息

Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Matunga, Mumbai 400019, India.

Department of Microbiology, Bombay Veterinary College, Parel, Mumbai 400012, India.

出版信息

Biosens Bioelectron. 2015 Aug 15;70:254-60. doi: 10.1016/j.bios.2015.03.045. Epub 2015 Mar 20.

Abstract

Presence of bacteria such as Brucella spp. in dairy products is an immense risk to public health. Point of care immunoassays are rapid in that they can quickly screen various samples in a relatively short amount of time, are sensitive, specific and offer a great advantage in accurate and fast diagnosis of infectious diseases. We have fabricated a point of care rapid diagnostic assay that employs fluorescent, micellar silica nanosensors capable of specifically detecting Brucella IgG antibodies in milk samples of afflicted animals. Currently, point of care detection assays are not commercially available for field testing of farm animals using milk samples. The nanosensing allows precise detection of antibodies with low sample volumes (50 μl). We demonstrate recognition of B. abortus antibodies through capture by fluorescent silica nanosensors using spiked and raw milk samples validated by ELISA and PCR. The test results are accurate and repeatable with high sensitivity and specificity, and a short assay time of 10 min for antigenic recognition and do not require any sample processing procedures such as isolation and separation. Additionally, well defined antigenic components and surface biomarkers of various disease causing microbes can be broadly incorporated within the purview of this technology for accurate and rapid detection of suspected bovine pathological conditions, and can largely enable rapid field testing that can be implemented in farms and food industry.

摘要

乳制品中存在布鲁氏菌等细菌对公众健康是一个巨大的威胁。即时检测免疫分析具有快速的特点,因为它们可以在相对较短的时间内快速筛选各种样本,具有灵敏度高、特异性强的优点,在传染病的准确快速诊断方面具有很大的优势。我们已经开发了一种即时检测快速诊断试剂盒,该试剂盒采用荧光胶束硅纳米传感器,能够特异性检测受感染动物牛奶样本中的布鲁氏菌 IgG 抗体。目前,即时检测检测试剂盒尚未商业化,无法用于现场检测农场动物的牛奶样本。纳米传感技术允许使用 50 μl 的小样本量进行精确的抗体检测。我们通过使用荧光硅纳米传感器对添加和原始牛奶样本进行 ELISA 和 PCR 验证,证明了对 B.abortus 抗体的识别。该测试具有高灵敏度和特异性,结果准确且可重复,抗原识别的检测时间短(10 分钟),并且不需要任何样品处理程序,如分离和分离。此外,各种致病微生物的明确抗原成分和表面生物标志物可以广泛纳入该技术的范围内,用于准确快速检测疑似牛的病理状况,并能够在农场和食品工业中实现广泛的快速现场测试。

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