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NOVEL METHOD FOR DETECTING PROTEIN OR PATHOGEN meetings

Foreign code F170008936
File No. (S2015-0897-N0)
Posted date Jan 19, 2017
Country WIPO
International application number 2016JP062081
International publication number WO 2016175049
Date of international filing Apr 15, 2016
Date of international publication Nov 3, 2016
Priority data
  • P2015-093132 (Apr 30, 2015) JP
Title NOVEL METHOD FOR DETECTING PROTEIN OR PATHOGEN meetings
Abstract Provided is a novel method for detecting a protein or a pathogen. The present invention pertains to a conductive diamond electrode wherein an element recognizing a protein or a pathogen is immobilized on the surface, and a method for detecting a protein or a pathogen using the conductive diamond electrode. More specifically, the present invention pertains to a diamond electrode wherein an element recognizing a virus or a protein thereof is immobilized on the surface, and a method for detecting a virus using the diamond electrode.
Outline of related art and contending technology BACKGROUND ART
Conventionally, the target pathogen, pathogenic bacteria, viruses and their respective proteins can be detected with high sensitivity have been desired. E.g. influenza virus (IFV) pandemics is the possibility, the detection of influenza virus quickly and accurately is demanded. IFV currently used as a method of detecting, using antibodies as the recognition device IFV immuno, using an array of sugar chains sugar chains detected, RT-PCR method using a gene, a hemagglutination assay using red blood cells and the like. However they are time and cost consuming, require expert knowledge or techniques.
Grabowska et al. using IFV hybridization detection methods are reported (Non-Patent Document 1). This method is via a thiol bond gold Au surface of the electrode, 2 is fixed to the two different oligonucleotide probes and the sensor is used, the hemagglutinin (HA) and neuraminidase (NA) both of the oligonucleotide target can be detected at the same time. Further Kamikawa et al., is covered with the electrical activity such as polyaniline, modified HA-H5N1 monoclonal antibody on the surface of the magnetic nanoparticles is used to report the detection IFV (Non-Patent Document 2). In this method, the virus H5N1 in interaction with the nanoparticles (MPC) in a strong magnetic force is recovered from the serum, the degree of electrochemical measurement is quantified by the virus.
Sakurai et al., fluorescence chromatography is used for rapid and sensitive of a seasonal flu typing was reported (Non-Patent Document 3). IFV clinical have been used in a method of detecting the detection sensitivity in the immunochromatography and 1000pfu, it is difficult to early detection of infection. Sakurai et al. in the publication, and a sensitizing agent of the virus antibody supplemented with labeled detection antibody with both the capture of the sandwich-type virus, densitometry analysis apparatus and fluorescence immunochromatography measuring device is further used is detected by using the conventional 100 times the sensitivity of the reported attained. However, a complementary oligonucleotide hybridization or antibody immobilized on the substrate, requires a huge manufacturing cost, the problem of storage stability.
Hassen et al., Electrochemical impedance spectroscopy was used to quantify the influenza A virus has been reported (Non-Patent Document 4). This document the antibody on the gold electrode surface to remove the sugar chain - - avidin fixing the thiol structure using devices, the influenza A virus is detected.
On the other hand the boron doped diamond electrode, glassy carbon or platinum electrodes and the like other conventional electrode materials of excellent and, in recent years, and have garnered attention. Can be a high thermal conductivity is much higher and hardness in addition to the well known properties of diamond, the boron doped diamond electrodes, large potential window, low background current, and the suction resistance is high, such as chemically inert to the attractive properties. In addition, the boron doped diamond electrode may be physically, chemically stable and excellent in durability.
Diamond electrode is provided with the sensor, an accurate determination of the catechol or catechol derivative is possible to perform the diamond electrode and the diamond sensor electrodes have been reported (Patent Document 1). This document also discloses that the modified diamond electrode 4 - pentenoic acid and oxalic acid is used which is the electrochemical detection.
Virus as well as various pathogenic bacteria, also for the pathogen, the antibody and the expensive device is not used, a high-sensitivity detection method has been desired.
Scope of claims (In Japanese)[請求項1]
タンパク質又は病原体を認識する素子が表面に固定化された導電性ダイヤモンド電極。
[請求項2]
タンパク質又は病原体を認識する素子が、インフルエンザウイルス、DNAウイルス、RNAウイルス、2本鎖DNAウイルス、1本鎖DNAウイルス、2本鎖RNAウイルス、1本鎖RNA(+)鎖ウイルス、1本鎖RNA(-)鎖ウイルス、1本鎖RNA逆転写ウイルス、2本鎖DNA逆転写ウイルス、ノロウイルス、ロタウイルス、風疹ウイルス、麻疹ウイルス、RSウイルス、ヘルペスウイルス、肝炎ウイルス、アデノウイルス、口蹄疫ウイルス、狂犬病ウイルス、ヒト免疫不全ウイルス、マイコプラズマ菌、ボツリヌス菌、百日咳菌、破傷風菌、ジフテリア菌、コレラ菌、赤痢菌、炭疽菌、病原性大腸菌、ブドウ球菌、サルモネラ菌、ウェルシュ菌又はセレウス菌を認識する素子である、請求項1に記載の電極。
[請求項3]
タンパク質又は病原体を認識する素子が、病原体タンパク質を認識するペプチドを有する、請求項1又は2に記載の電極。
[請求項4]
タンパク質又は病原体を認識する素子が、インフルエンザウイルスのヘマグルチニンタンパク質(HA)、インフルエンザウイルスのノイラミニダーゼ(NA)、M1タンパク質若しくはM2タンパク質、マイコプラズマ細菌のP1タンパク質、膜抗原タンパク質若しくはリボソームタンパク質L7/L12、ボツリヌス毒素、百日咳毒素、破傷風菌毒素、ジフテリア毒素、ウェルシュ菌のα毒素、コレラ菌毒素、ベロ毒素、炭疽菌毒素、大腸菌由来エンテロトキシン、ブドウ球菌由来エンテロトキシン、サルモネラ菌由来エンテロトキシン又はセレウス菌由来エンテロトキシンを認識するペプチドを有する、請求項1~3のいずれか1項に記載の電極。
[請求項5]
タンパク質又は病原体を認識する素子が、タンパク質又は病原体を認識する分子及びリンカー部分を有し、該タンパク質又は病原体を認識する分子が該リンカー部分を介してダイヤモンド電極の表面に固定化されている、請求項1~4のいずれか1項に記載の電極。
[請求項6]
請求項1~5のいずれか1項に記載の電極を備えた、タンパク質又は病原体を検出する装置。
[請求項7]
請求項1~5のいずれか1項に記載の電極又は請求項6に記載の装置を用いた、タンパク質又は病原体を検出する方法。
[請求項8]
サイクリックボルタンメトリー測定又は電気化学インピーダンス測定によりタンパク質又は病原体を検出する、請求項7に記載の検出方法。
[請求項9]
導電性ダイヤモンド電極の表面に、タンパク質又は病原体を認識する分子を固定化してタンパク質又は病原体を認識する素子を形成する工程を含む、タンパク質又は病原体検出用導電性ダイヤモンド電極の製造方法。
[請求項10]
リンカー分子をダイヤモンド電極の表面に固定化する工程、及びリンカー分子とタンパク質又は病原体を認識する分子とを連結する工程を含む、請求項9に記載の製造方法。
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • KEIO UNIVERSITY
  • Inventor
  • SATO Toshinori
  • MATSUBARA Teruhiko
  • EINAGA Yasuaki
  • YAMAMOTO Takashi
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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