title: Ultrasensitive Detection of Ebola Virus Oligonucleotide Based on Upconversion Nanoprobe/Nanoporous Membrane System.
abstract: Ebola outbreaks are currently of great concern, and therefore, development of effective diagnosis methods is urgently needed. The key for lethal virus detection is high sensitivity, since early-stage detection of virus may increase the probability of survival. Here, we propose a luminescence scheme of assay consisting of BaGdF5:Yb/Er upconversion nanoparticles (UCNPs) conjugated with oligonucleotide probe and gold nanoparticles (AuNPs) linked with target Ebola virus oligonucleotide. As a proof of concept, a homogeneous assay was fabricated and tested, yielding a detection limit at picomolar level. The luminescence resonance energy transfer is ascribed to the spectral overlapping of upconversion luminescence and the absorption characteristics of AuNPs. Moreover, we anchored the UCNPs and AuNPs on a nanoporous alumina (NAAO) membrane to form a heterogeneous assay. Importantly, the detection limit was greatly improved, exhibiting a remarkable value at the femtomolar level. The enhancement is attributed to the increased light-matter interaction throughout the nanopore walls of the NAAO membrane. The specificity test suggested that the nanoprobes were specific to Ebola virus oligonucleotides. The strategy combining UCNPs, AuNPs, and NAAO membrane provides new insight into low-cost, rapid, and ultrasensitive detection of different diseases. Furthermore, we explored the feasibility of clinical application by using inactivated Ebola virus samples. The detection results showed great potential of our heterogeneous design for practical application.
authors.affiliation: Department of Applied Physics, The Hong Kong Polytechnic University , Hung Hom, Kowloon, Hong Kong, P. R. China.
authors.forename: Ming-Kiu
authors.initials: MK
authors.lastname: Tsang
authors.affiliation: Interdisciplinary Division of Biomedical Engineering, The Hong Kong Polytechnic University , Hung Hom, Kowloon, Hong Kong, P. R. China.|Institute of Ocean Research, Zhejiang University of Technology , Hangzhou, Zhejiang 310014, P. R. China.
authors.forename: WeiWei
authors.initials: W
authors.lastname: Ye
authors.affiliation: National Center for Clinical Laboratory, Beijing Hospital of the Ministry of Health , No. 1 Dahua Road, Dongdan, Beijing 100730, P. R. China.
authors.forename: Guojing
authors.initials: G
authors.lastname: Wang
authors.affiliation: National Center for Clinical Laboratory, Beijing Hospital of the Ministry of Health , No. 1 Dahua Road, Dongdan, Beijing 100730, P. R. China.
authors.forename: Jingming
authors.initials: J
authors.lastname: Li
authors.affiliation: Interdisciplinary Division of Biomedical Engineering, The Hong Kong Polytechnic University , Hung Hom, Kowloon, Hong Kong, P. R. China.
authors.forename: Mo
authors.initials: M
authors.lastname: Yang
authors.affiliation: Department of Applied Physics, The Hong Kong Polytechnic University , Hung Hom, Kowloon, Hong Kong, P. R. China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Hao
mesh_terms: D017609:Barium Compounds / Q000737:chemistry*; D001681:Biological Assay; D029043:Ebolavirus / Q000235:genetics / Q000302:isolation & purification*; D005063:Europium / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D005459:Fluorides; D006046:Gold / Q000737:chemistry*; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis / Q000821:virology; D006801:Humans; D057230:Limit of Detection; D008567:Membranes, Artificial; D053768:Metal Nanoparticles / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D016062:Porosity; D015018:Ytterbium / Q000737:chemistry
chemical_list: D017609:Barium Compounds; D008567:Membranes, Artificial; D009841:Oligonucleotides; D005063:Europium; D006046:Gold; D015018:Ytterbium; D005459:Fluorides
keywords: Ebola virus RNA; LRET; biosensor; nanoprobe; upconversion