A novel carbon nanoparticle probe-based ultrasensitive lateral flow assay for rapid detection of Ebola virus.

Yanqiu Wei; Yongcheng Duan; Yuhai BI; Meng Wang; Yunlong LI; Xuan Wang; Wei LI; Wenhui Fan; Jing Wang; Wenjun Liu; Limin Yang

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A novel carbon nanoparticle probe-based ultrasensitive lateral flow assay for rapid detection of Ebola virus.

Author: Yanqiu WEI ¹ ; Yongcheng DUAN ² ; Yuhai BI ¹ ; Meng WANG ¹ ; Yunlong LI ¹ ; Xuan WANG ¹ ; Wei LI ³ ; Wenhui FAN ¹ ; Jing WANG ⁴ ; Wenjun LIU ¹ ; Limin YANG ¹
Author Information

1. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
2. College of Animal Science and Technology, Guangxi University, Nanning 530004, Guangxi, China.
3. Goodwe Biotechnology Taizhou Co., Ltd., Taizhou 225300, Jiangsu, China.
4. National Institute of Metrology, Beijing 100029, China.
Publication Type:Journal Article
Keywords: Ebola virus; POC; colloidal carbon; lateral flow immunochromatography; matrix protein; monoclonal antibodies
MeSH: Animals; Carbon; Ebolavirus; Hemorrhagic Fever, Ebola; Humans; Nanoparticles; Rabbits
From: Chinese Journal of Biotechnology 2018;34(12):2025-2034
CountryChina
Language:Chinese
Abstract: Ebola virus (EBOV) is an extremely contagious pathogen first discovered in Africa associated with severe hemorrhagic disease in humans and nonhuman primates, which has resulted in at least 28 500 suspected cases and 11 300 confirmed deaths in 2014-2016 Ebola epidemic in West Africa. Rapid and sensitive detection of EBOV is the key to increasing the probability of survival and reducing infection rates in pandemic regions. Here, we report an ultrasensitive and instrument-free EBOV detection assay based on colloidal carbon immunochromatography. Carbon nanoparticle-labeled rabbit anti-EBOV-VP40 IgG were concentrated in the conjugate pad, monoclonal antibody (McAb, 4B7F9) against EBOV-VP40 and goat anti-rabbit IgG were immobilized on the nitrocellulose membrane with 2 μL/cm at a concentration of 1 mg/mL as test and control lines, respectively. Then the sample application pad, conjugate release pad, nitrocellulose membrane and absorbent pad were assembled into a lateral flow test strip. The test strip shows strong specificity against related viruses that share similar clinical symptoms and geographic range with EBOV, including marburg virus, influenza virus, yellow fever virus and dengue virus. In addition, 1 500 negative serums were tested with false-positive rate of 1.3‰ which significantly lower than that of ReEBOV™ colloidal gold test kit recommended by World Health Organization (WHO). The sensitivity of this strip was analyzed using inactivated EBOV with detection limit of 100 ng/mL (10⁶ copies/mL) which clearly higher than that of ReEBOV™ dipstick (10⁸ copies/mL). Furthermore, the strip showed excellent thermal stability characteristics in room temperature and could be as a point-of-care (POC), ultra-sensitive and specific promising candidate for EBOV serological screening in rural Africa or entry/exit ports.