Fast and Parallel Detection of Four Ebola Virus Species on a Microfluidic-Chip-Based Portable Reverse Transcription Loop-Mediated Isothermal Amplification System.

Xue Lin,Kai Jiang,Xiangyu Jin,Rongxin Fu,Han Yang,Guoliang Huang,Ya Su,Bin Xu,Yong Guo,Ying Lu,Ruliang Wang

doi:10.3390/mi10110777

Xue Lin, Kai Jiang + Show 9 more

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https://doi.org/10.3390/mi10110777

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Journal: Micromachines	Publication Date: Nov 14, 2019
Citations: 19	License type: CC BY 4.0

Affiliation: Tsinghua University

Abstract

Considering the lack of official vaccines and medicines for Ebola virus infection, reliable diagnostic methods are necessary for the control of the outbreak and the spread of the disease. We developed a microfluidic-chip-based portable system for fast and parallel detection of four Ebola virus species. The system is based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and consists of four specific LAMP primers, a disc microfluidic chip, and a portable real-time fluorescence detector. It could specifically and parallelly distinguish four species of the Ebola virus after only one sampling, including the Zaire Ebola virus, the Sudan Ebola virus, the Bundibugyo Ebola virus, and the Tai Forest Ebola virus, without cross-contamination. The limit of detection was as small as 10 copies per reaction, while the total consumption of sample and reagent was 0.94 μL per reaction. The final results could be obtained in 50 min after one addition of sample and reagent mixture. This approach provides simplicity, high sensitivity, and multi-target parallel detection at a low cost, which could enable convenient and effective on-site detections of the Ebola virus in the outdoors, remote areas, and modern hospitals.

Highlights

Ebola virus is a filovirus containing a non-segmented, negative-sense, and single-stranded ribonucleic acid (RNA) genome and belongs to the family Filoviridae [1,2]
We developed a microfluidic-chip-based portable system for fast and parallel detection of the four Ebola virus species
By using the developed portable real-time fluorescence detector and a disc microfluidic chip, 24 independent nucleic acid amplifications could be simultaneously performed on a single chip after a single sample loading

Summary

Introduction

Ebola virus is a filovirus containing a non-segmented, negative-sense, and single-stranded ribonucleic acid (RNA) genome and belongs to the family Filoviridae [1,2]. A new outbreak was declared in February 2019 in the towns of Butembo and Katwa in the Northeast area of the Democratic Republic of Congo. As of 15 July 2019, a total of 2512 cases were reported, including 2418 confirmed and 94 probable cases, of which 1676 died (overall fatality ratio: 67%). The WHO declared the Ebola virus disease in the Democratic Republic of Congo as a public health emergency of international concern on 17 July [5]. The prevention and the control of the Ebola virus is still a serious worldwide issue. No proven vaccines or treatments for Ebola have been reported [4]. It is crucial to develop Ebola virus specific detection and fast diagnostic method to manage the spread of the disease on site

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