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通过微流控实时聚合酶链反应系统快速、多重且自动化检测眼内炎中的细菌和真菌。

Rapid, multiplex and automated detection of bacteria and fungi in endophthalmitis via a microfluidic real-time pcr system.

作者信息

Wang Siyu, Liu Yiteng, Li Yingqi, Gao Yibo, Zou Zhongliang, Xu Na, Song Qi, Liu Fangyan, Song Yihong, Wang Xian, Fan Zixin

机构信息

Department of Ophthalmology, the First Affiliated Hospital of China Medical University, No. 155 Nanjing Bei Street, Heping District, Shenyang City, 110001, Liaoning Province, PR China.

Division of Emerging Interdisciplinary Areas, Academy of Interdisciplinary Studies, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, 999077, Hong Kong SAR, China.

出版信息

J Ophthalmic Inflamm Infect. 2024 Dec 18;14(1):64. doi: 10.1186/s12348-024-00446-6.

DOI:10.1186/s12348-024-00446-6
PMID:39694980
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11655742/
Abstract

BACKGROUND

Endophthalmitis is an ophthalmologic emergency requiring accurate and rapid diagnosis for treatment. Currently, the diagnosis commonly relies on culture and molecular biology, which falls short of clinical rapid diagnosis. The purpose of this study was to evaluate the feasibility of a self-build Microfluidic Real-time Polymerase Chain Reaction (RT-PCR) System for rapidly identifying potential pathogens of endophthalmitis.

METHODS

This study included 22 patients who presented to Shenzhen Eye Hospital and the Ophthalmology Department of the Affiliated Hospital of Guizhou Medical University in China between January 2023 and March 2024. The samples were cultured using conventional methods and underwent Microfluidic RT-PCR and metagenomic next-generation sequencing (mNGS).

RESULTS

The Microfluidic RT-PCR System identified pathogens in 11 of 22 cases (50.00%), compared with 40.91% for microbiology culture. 14 cases (63.64%) had concordant results, and 5 cases were positive for the microfluidic system only. The agreements between culture and microfluidic system, as well as culture and mNGS were 100.00% (6/6) and 50.00% (3/6), respectively. The average waiting time for the microfluidic system was about 30 min if excepting DNA extraction time, which was much shorter than 2.88 days for culture and 1.57 days for mNGS.

CONCLUSION

The microfluidic-based RT-PCR system was preliminarily proved to be a sensitive, easy-to-operate, and rapid in-hospital technology. It is expected to become a rapid diagnostic platform for endophthalmitis.

摘要

背景

眼内炎是一种眼科急症，需要准确快速的诊断以进行治疗。目前，诊断通常依赖于培养和分子生物学方法，但这难以满足临床快速诊断的需求。本研究的目的是评估自行构建的微流控实时聚合酶链反应（RT-PCR）系统用于快速鉴定眼内炎潜在病原体的可行性。

方法

本研究纳入了2023年1月至2024年3月期间在中国深圳眼科医院和贵州医科大学附属医院眼科就诊的22例患者。样本采用传统方法进行培养，并进行微流控RT-PCR和宏基因组下一代测序（mNGS）。

结果

微流控RT-PCR系统在22例中的11例（50.00%）中鉴定出病原体，而微生物培养的阳性率为40.91%。14例（63.64%）结果一致，5例仅微流控系统呈阳性。培养与微流控系统以及培养与mNGS之间的一致性分别为100.00%（6/6）和50.00%（3/6）。如果不包括DNA提取时间，微流控系统的平均等待时间约为30分钟，远短于培养的2.88天和mNGS的1.57天。

结论

基于微流控的RT-PCR系统初步证明是一种敏感、易于操作且快速的院内技术。有望成为眼内炎的快速诊断平台。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/a0bacbdf9528/12348_2024_446_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/1362c5273a0d/12348_2024_446_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/47345b2ec0c8/12348_2024_446_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/9fc2abb3c5fd/12348_2024_446_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/e39f6afbfc34/12348_2024_446_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/a0bacbdf9528/12348_2024_446_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/1362c5273a0d/12348_2024_446_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/47345b2ec0c8/12348_2024_446_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/9fc2abb3c5fd/12348_2024_446_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/e39f6afbfc34/12348_2024_446_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7365/11655742/a0bacbdf9528/12348_2024_446_Fig3_HTML.jpg

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Biosensors (Basel). 2023 Mar 21;13(3):410. doi: 10.3390/bios13030410.
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Lower fluidic resistance of double-layer droplet continuous flow PCR microfluidic chip for rapid detection of bacteria.双层液滴连续流 PCR 微流控芯片具有较低的流动阻力，可快速检测细菌。
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