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Naked-Eye Enumeration of Single Chlamydia pneumoniae Based on Light Scattering of Gold Nanoparticle Probe.
ACS Sensors ( IF 8.2 ) Pub Date : 2020-03-24 , DOI: 10.1021/acssensors.0c00150 Fenglei Chen 1 , Tao Di 1 , Chih-Tsung Yang 2 , Tianyu Zhang 1 , Benjamin Thierry 2 , Xin Zhou 1
ACS Sensors ( IF 8.2 ) Pub Date : 2020-03-24 , DOI: 10.1021/acssensors.0c00150 Fenglei Chen 1 , Tao Di 1 , Chih-Tsung Yang 2 , Tianyu Zhang 1 , Benjamin Thierry 2 , Xin Zhou 1
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Chlamydia pneumoniae is a spherical zoonotic pathogen with a diameter of ∼200 nm, which can lead to a wide range of acute and chronic diseases in human body. Early and reliable on-site detection of C. pneumoniae is the key step to control the spread of the pathogen. However, the lack of a current technology with advantages of rapidity, ultrasensitivity, and convenience limits the implementation of traditional techniques for on-site detection of C. pneumoniae. Herein, we developed a naked-eye counting of C. pneumoniae based on the light scattering properties of gold nanoparticle (GNP) under dark-field microscopy (termed "GNP-labeled dark-field counting strategy"). The recognition of single C. pneumoniae by anti-C. pneumoniae antibodies-functionalized GNP probes with size of 15 nm leads to the formation of wreath-like structure due to the strong scattered light resulted from hundreds of GNP probes binding on one C. pneumoniae under dark-field microscopy. Hundreds of GNP probes can bind to the surface of C. pneumoniae due to the high stability and specificity of the nucleic acid immuno-GNP probes, which generates by the hybridization of DNA-modified GNP with DNA-functionalized antibodies. The limit of detection (LOD) of the GNP-labeled dark-field counting strategy for C. pneumoniae detection in spiked samples or real samples is down to four C. pneumoniae per microliter, which is about 4 times more sensitive than that of quantitative polymerase chain reaction (qPCR). Together with the advantages of the strong light scattering characteristic of aggregated GNPs under dark-field microscopy and the specific identification of functionalized GNP probes, we can detect C. pneumoniae in less than 30 min using a cheap and portable microscope even if the sample contains only a few targets of interest and other species at high concentration. The GNP-labeled dark-field counting strategy meets the demands of rapid detection, low cost, easy to operate, and on-site detection, which paves the way for early and on-site detection of infectious pathogens.
中文翻译：
基于金纳米粒子探针的光散射的单眼肺炎衣原体裸眼计数。
肺炎衣原体是一种球形的人畜共患病原体，直径约为200 nm，可导致多种人体急性和慢性疾病。早期可靠的肺炎衣原体现场检测是控制病原体传播的关键步骤。然而，缺乏具有快速，超灵敏和方便的优点的当前技术限制了用于现场检测肺炎衣原体的传统技术的实施。在此，我们根据金纳米颗粒（GNP）在暗场显微镜下的光散射特性（称为“ GNP标记的暗场计数策略”）开发了肉眼计数的肺炎衣原体。抗C对单个肺炎衣原体的识别。直径为15 nm的肺炎球菌抗体功能化GNP探针由于在暗场显微镜下结合在一个肺炎衣原体上的数百个GNP探针产生的强散射光而导致形成花圈状结构。由于核酸免疫GNP探针的高度稳定性和特异性，数百种GNP探针可以结合到肺炎衣原体的表面，该核酸免疫GNP探针是通过将DNA修饰的GNP与DNA功能化的抗体杂交而产生的。GNP标记的暗场计数策略在加标样品或实际样品中检测肺炎衣原体的检测下限（LOD）降至每微升4个肺炎衣原体，比定量聚合酶的敏感性高约4倍连锁反应（qPCR）。加上在暗场显微镜下聚集的GNP的强光散射特性和功能化GNP探针的特异性鉴定的优点，即使样品中仅含有样品，我们也可以使用廉价的便携式显微镜在不到30分钟的时间内检测出肺炎衣原体。一些感兴趣的目标和其他高浓度物种。GNP标记的暗场计数策略可满足快速检测，低成本，易于操作和现场检测的需求，这为传染病病原体的早期和现场检测铺平了道路。
更新日期：2020-03-24
中文翻译：
基于金纳米粒子探针的光散射的单眼肺炎衣原体裸眼计数。
肺炎衣原体是一种球形的人畜共患病原体，直径约为200 nm，可导致多种人体急性和慢性疾病。早期可靠的肺炎衣原体现场检测是控制病原体传播的关键步骤。然而，缺乏具有快速，超灵敏和方便的优点的当前技术限制了用于现场检测肺炎衣原体的传统技术的实施。在此，我们根据金纳米颗粒（GNP）在暗场显微镜下的光散射特性（称为“ GNP标记的暗场计数策略”）开发了肉眼计数的肺炎衣原体。抗C对单个肺炎衣原体的识别。直径为15 nm的肺炎球菌抗体功能化GNP探针由于在暗场显微镜下结合在一个肺炎衣原体上的数百个GNP探针产生的强散射光而导致形成花圈状结构。由于核酸免疫GNP探针的高度稳定性和特异性，数百种GNP探针可以结合到肺炎衣原体的表面，该核酸免疫GNP探针是通过将DNA修饰的GNP与DNA功能化的抗体杂交而产生的。GNP标记的暗场计数策略在加标样品或实际样品中检测肺炎衣原体的检测下限（LOD）降至每微升4个肺炎衣原体，比定量聚合酶的敏感性高约4倍连锁反应（qPCR）。加上在暗场显微镜下聚集的GNP的强光散射特性和功能化GNP探针的特异性鉴定的优点，即使样品中仅含有样品，我们也可以使用廉价的便携式显微镜在不到30分钟的时间内检测出肺炎衣原体。一些感兴趣的目标和其他高浓度物种。GNP标记的暗场计数策略可满足快速检测，低成本，易于操作和现场检测的需求，这为传染病病原体的早期和现场检测铺平了道路。
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