Identification of pathogenic bacteria in complex samples using a smartphone based fluorescence microscope.
Identification of pathogenic bacteria in complex samples using a smartphone based fluorescence microscope.
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DOI：
10.1039/c8ra06473c
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发表时间：
2018-10-26
期刊：
影响因子：
3.9
通讯作者：
Westerlund, Fredrik
中科院分区：
文献类型：
--
作者：
Mueller, Vilhelm;Sousa, Jose M.;Koydemir, Hatice Ceylan;Veli, Muhammed;Tseng, Derek;Cerqueira, Laura;Ozcan, Aydogan;Azevedo, Nuno F.;Westerlund, Fredrik
基于生物分子的荧光成像的诊断通常在装备精良的实验室中进行，并且通常不适合于远程和资源有限的环境。在这里，我们展示了一种紧凑，轻便和具有成本效益的基于智能手机的荧光显微镜的开发，能够检测荧光标记细菌的信号。通过优化基于肽核酸（PNA）的荧光原位杂交（FISH）检测，我们展示了使用基于智能手机的显微镜快速鉴定病原菌。我们评估了使用一般的核酸染色以及物种特异性PNA探针，并证明了移动的平台可以检测细菌的灵敏度与传统的荧光显微镜。基于PNA的FISH检测与基于智能手机的荧光显微镜相结合，使我们能够在培养前以低至每30 g PIF 10 CFU的初始浓度定性分析受污染的婴儿配方奶粉（PIF）中的致病菌。重要的是，检测可以直接在智能手机屏幕上完成，无需额外的图像分析。该测定法应简单明了，以适应临床样品中的细菌鉴定。该平台的成本效益、现场便携性和简单性将为其在资源有限的环境和护理点办公室中的使用创造各种机会，从而开辟了基于其他荧光诊断测定的无数其他应用。一种基于智能手机的荧光显微镜，用于使用物种特异性肽核酸探针检测复杂样品中的细菌。
Diagnostics based on fluorescence imaging of biomolecules is typically performed in well-equipped laboratories and is in general not suitable for remote and resource limited settings. Here we demonstrate the development of a compact, lightweight and cost-effective smartphone-based fluorescence microscope, capable of detecting signals from fluorescently labeled bacteria. By optimizing a peptide nucleic acid (PNA) based fluorescence in situ hybridization (FISH) assay, we demonstrate the use of the smartphone-based microscope for rapid identification of pathogenic bacteria. We evaluated the use of both a general nucleic acid stain as well as species-specific PNA probes and demonstrated that the mobile platform can detect bacteria with a sensitivity comparable to that of a conventional fluorescence microscope. The PNA-based FISH assay, in combination with the smartphone-based fluorescence microscope, allowed us to qualitatively analyze pathogenic bacteria in contaminated powdered infant formula (PIF) at initial concentrations prior to cultivation as low as 10 CFU per 30 g of PIF. Importantly, the detection can be done directly on the smartphone screen, without the need for additional image analysis. The assay should be straightforward to adapt for bacterial identification also in clinical samples. The cost-effectiveness, field-portability and simplicity of this platform will create various opportunities for its use in resource limited settings and point-of-care offices, opening up a myriad of additional applications based on other fluorescence-based diagnostic assays. A smartphone-based fluorescence microscope for detection of bacteria in complex samples using a species-specific peptide nucleic acid probe.
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影响因子：
5.6
作者：
Cerqueira, Laura;Azevedo, Nuno F.;Almeida, Carina;Jardim, Tatiana;Keevil, Charles William;Vieira, Maria J.
通讯作者：
Vieira, Maria J.
影响因子：
3.3
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
Venkitanarayanan, KS
影响因子：
10.4
作者：
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通讯作者：
Ozcan, Aydogan
影响因子：
2.2
作者：
Liu, Y;Cai, XN;Huang, XT
通讯作者：
Huang, XT