JP4264469B2 - Testing method for dementia - Google Patents
Testing method for dementia Download PDFInfo
- Publication number
- JP4264469B2 JP4264469B2 JP2003043452A JP2003043452A JP4264469B2 JP 4264469 B2 JP4264469 B2 JP 4264469B2 JP 2003043452 A JP2003043452 A JP 2003043452A JP 2003043452 A JP2003043452 A JP 2003043452A JP 4264469 B2 JP4264469 B2 JP 4264469B2
- Authority
- JP
- Japan
- Prior art keywords
- annexin
- dementia
- antibody
- blood
- concentration
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 206010012289 Dementia Diseases 0.000 title claims description 21
- 238000012360 testing method Methods 0.000 title claims description 13
- 102000004121 Annexin A5 Human genes 0.000 claims description 30
- 108090000672 Annexin A5 Proteins 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 20
- 210000004369 blood Anatomy 0.000 claims description 14
- 239000008280 blood Substances 0.000 claims description 14
- 208000024827 Alzheimer disease Diseases 0.000 description 12
- 102000004148 Annexin A4 Human genes 0.000 description 8
- 108090000669 Annexin A4 Proteins 0.000 description 8
- 238000005259 measurement Methods 0.000 description 8
- 210000002381 plasma Anatomy 0.000 description 8
- 102000000412 Annexin Human genes 0.000 description 6
- 108050008874 Annexin Proteins 0.000 description 6
- 238000002965 ELISA Methods 0.000 description 5
- 239000007790 solid phase Substances 0.000 description 5
- 238000010998 test method Methods 0.000 description 5
- 208000037259 Amyloid Plaque Diseases 0.000 description 4
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 4
- 238000001262 western blot Methods 0.000 description 4
- 208000007848 Alcoholism Diseases 0.000 description 3
- 201000007930 alcohol dependence Diseases 0.000 description 3
- 210000001124 body fluid Anatomy 0.000 description 3
- 239000010839 body fluid Substances 0.000 description 3
- 210000004556 brain Anatomy 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000003317 immunochromatography Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 239000004816 latex Substances 0.000 description 2
- 229920000126 latex Polymers 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 102000013455 Amyloid beta-Peptides Human genes 0.000 description 1
- 108010090849 Amyloid beta-Peptides Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 125000003158 alcohol group Chemical group 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 210000001130 astrocyte Anatomy 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000003710 cerebral cortex Anatomy 0.000 description 1
- 208000013677 cerebrovascular dementia Diseases 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 208000010877 cognitive disease Diseases 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000003412 degenerative effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 108091005899 fibrous proteins Proteins 0.000 description 1
- 102000034240 fibrous proteins Human genes 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000001114 immunoprecipitation Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 1
- 210000002241 neurite Anatomy 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 210000002826 placenta Anatomy 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Description
【０００１】
【発明の属する技術分野】
本発明は臨床検査の分野で用いられ、とりわけ痴呆症の検査方法として用いられる。
【０００２】
【従来の技術】
アルツハイマー病（ＡＤ）はおもに老年期に発病する進行性の神経変性疾患で認知機能障害を特徴とする。ＡＤにおける痴呆症の程度はＡＤ患者の大脳皮質に見出される老人班（ＳＰ）の数に関連づけられてきた (例えば非特許文献１)。そして組織学的には、ＳＰは退化性神経突起及び活性の星状膠細胞に囲まれ、また生化学的には、ＳＰはβアミロイドと呼ばれる小分子の不溶性繊維状タンパク質を含んでいる(例えば非特許文献２) 。ＡＤの原因は不明であり、ＡＤの治療法も乏しいのが現状である。ＡＤの診断は診断基準によるところの除外診断および脳の画像診断を主とし、それ以外の客観的方法はほとんどない。すなわち、発症前或いは発症後の患者におけるＡＤ鑑別診断のための簡便な検査法は存在しない。特許文献１によれば脳脊髄液中の分子量３３０００の蛋白質を免疫学的に測定する方法が開示されているが、侵襲性の高い脳脊髄液を用いなければならないといった問題がある。脳血管障害及びＡＤ患者の脳において数種のアネキシンが多量に発現している報告もある（例えば、非特許文献３）。さらにアルコール依存症の患者の脳にはアネキシンＩＶが正常群よりも多く発現していることも報告されている（非特許文献４）。以上の例はすべて死後患者から採取された脳脊髄液や脳組織の分析が行われているが、これらは診断治療に用いることができる方法ではない。非特許文献４に示されているように、アルコール依存症患者において血中の抗アネキシンＩＶ抗体量がアルコール依存症の診断に有効であるという報告は、簡便な検査方法といえる。しかしながら、前にも述べたようにＡＤの簡便な診断検査の方法は実用化されておらず、侵襲性が低くかつ簡単に患者より採取できる血液等の体液を用いるＡＤの簡便な検査方法の開発が望まれている。
【０００３】
【特許文献１】
特開平７−２５３４２９号。
【特許文献２】
特開平７−７２１４７号。
【非特許文献１】
Nature、1966、209巻、１０９頁。
【非特許文献２】
Biochem Biophys
Res Commun. 120巻、885-890頁（１９８４）
【非特許文献３】
Am J Pathol,
145, 640-649
(1994)
【非特許文献４】
Alcohol Clin
Exp Res, 25, 55S-58S (2001)
【０００４】
【発明が解決しようとする課題】
本発明の課題は痴呆症の検査方法であって、血液等の簡単に採取できる体液を用いて簡便に検査できる方法を提供することである。
【０００５】
【課題を決するための手段】
本発明者らは、痴呆症の簡便な検査方法を鋭意研究した結果、血液中のアネキシン類、とりわけアネキシンＶの濃度が痴呆の発症と深く関連することから簡便な痴呆症の検査が可能であることを見出し、本発明を完成させた。
【０００６】
本発明は以下の構成からなる。
１、血液中のアネキシンＶ濃度を測定することを特徴とする痴呆症の検査方法。
２、血液が血漿である前記１に記載の方法。
３、前記１〜２に記載の方法を用いる試薬又はキット。
【０００７】
【発明の実施の形態】
本発明は、痴呆症の検査に関するものであり、体液中のアネキシン類、とりわけ
アネキシンＶの濃度を測定することによる痴呆症の検査方法に関するものである。本発明は血液、より好ましくは血漿中のアネキシンＶを特異的な抗体（抗体全分子又は抗体活性を有する部分断片を含む）を用いて免疫学的に測定することにより実施される。用いる抗体は、ポリクローナル抗体及びモノクローナル抗体のいずれも用いることが可能であり、特に限定されるものではない。免疫学的測定法は、ＥＩＡ、ＲＩＡ、ラテックス凝集反応、免疫沈降法、ウエスタンブロット、ドットブロット、イムノクロマト法等公知の方法が適用される。アネキシンＶ測定のためのＥＩＡ法を例にとれば、固相に固定化された被検試料中のアネキシンＶを反応させ、さらに酵素標識した抗アネキシンＶ抗体を反応させ、固相に捕獲された標識抗体の量を検出することにより試料中のアネキシンＶの濃度を測定することができる。
【０００８】
またアネキシンＶの生物学的性質を利用して、例えばリン脂質、好ましくはホスファチジルセリンを固定化した固相に被検試料中のアネキシンＶを捕獲した後、標識抗アネキシンＶ抗体を反応させ、その反応量を検出することによっても測定できる。
【０００９】
本発明は、血液中のアネキシンＶと他のアネキシン類、例えばアネキシンＩＶの濃度を組み合わせて検査してもよい。例えば両者の比を算出すること、両方の測定値について、それぞれ閾値を設定しておき、その閾値を両方が超える、どちらか片方が超える、どちらも超えない等の判定結果から、その診断的有用性を検討することにより行われる。さらにアネキシンＶとアネキシンＩＶの測定は、血液中のアネキシンＶが測定されていれば、必ずしも同一被検試料である必要はなく、例えば血清と血漿、脳脊髄液と血漿等の異なる試料の組合せでも可能である。
【００１０】
アネキシン分子種は、痴呆やアルコール依存症などの神経精神疾患で発現変化することが報告されている（例えば、非特許文献２、３）。前述のようにアネキシンＶとアネキシンＩＶを組み合わせて用いることにより、アネキシンＶ濃度の上昇が痴呆症によるものか、あるいは他の疾患によるものかを的確に鑑別できる。逆にアネキシンＩＶの上昇がアルコール依存性か否かを検証するのにもアネキシンＶとの組み合わせが有効である。以上のように、アネキシンＶ濃度及びアネキシンＩＶ濃度を組み合わせて検査することにより特異的な痴呆症の検査が期待できる。
【００１１】
アネキシンＶとアネキシンＩＶの測定を組み合わせて行う方法は、前述のようにそれぞれ個々に構築された免疫学的手法、例えばＥＩＡ、ラテックス比濁法等を用いてそれぞれの濃度を測定することはもちろんのことであるが、それ以外にも、それぞれの抗体（そのフラグメントを含む）をひとつの反応容器に固定化しておき、そして被検試料を反応させ、ひとつの反応容器内にアネキシンＶとアネキシンＩＶの2種類の分子を捕獲し、それらを、それぞれの分子に対する抗体（そのフラグメントを含む）を異なる標識物質で標識した標識抗体で検出することによる方法も可能である。さらにはイムノクロマト法を利用して、同時に2種類の抗原を検出することも可能である（図１）。
【００１２】
【実施例】
本発明の実施例を以下に示すが、本発明は実施例に限定されるものではない。
【実施例１】
（アネキシンＶ濃度測定用ＥＬＩＳＡの構築）
ヒト胎盤より精製したアネキシンＶを免疫原としてマウスモノクローナル抗体を調製した。調製したモノクローナル抗体（クローンNo.２３）を９６穴のＥＬＩＳＡ用マイクロタイタープレートに固相化し、抗アネキシンＶ抗体固相プレートを調製した。固相プレートに用いたクローンと異なるモノクローナル抗体（クローンNo.４９）をビオチン標識して標識抗体を調製した。これらを組み合わせてアネキシンＶ測定用ＥＬＩＳＡ試薬を構築した。
【００１３】
【実施例２】
（ウエスタンブロットでの検討）
痴呆患者11名（アルツハイマー病患者：男性3名、女性6名、脳血管性痴呆患者：女性2名）及びコントロール10名（男性5名、女性5名）の血漿中のアネキシンをウエスタンブロットで検出した。各患者は予め長谷川式簡易知能評価スケールにて知能得点を調べた。各血漿１ｍＬをＰＢＳ緩衝液で10倍に希釈し、ホスファチジルセリンとホスファチジルコリンの混合比を１：９（モル比）のリポソーム１００μｇを加え、さらに終濃度で６ｍＭ塩化カルシウムを加え15分間インキュベートした。その後遠心操作により沈殿画分を集め、洗浄後ＳＤＳ−ＰＡＧＥを行い、ＰＶＤＦ膜に転写して、ウサギ由来の抗アネキシンＶ抗体でウエスタンブロットを行い、血液中のアネキシンＶの存在と痴呆の関連を調べた。コントロール群ではアネキシンＶのいずれも検出されなかったのに対して、痴呆症ではアネキシンＶは１１例中１０例（感度９０．９％）において検出され血液中のアネキシンＶの検出又は測定が痴呆症の検査に有用であることがわかった。
【００１４】
【実施例３】
（酵素免疫測定法での検討）
実施例２で検討した検体中のアネキシンＶ濃度を実施例１で調製したＥＬＩＳＡ法で測定した。その結果を表１に示した。
【００１５】
【表１】
ＥＬＩＳＡ法で測定において、痴呆症の患者血漿中のアネキシンＶ濃度はコントロール群に比べて有意に高い値を示し、血漿中のアネキシンＶ濃度の測定が痴呆症の検査に有用であることがわかった。
【００１７】
【発明の効果】
本発明は、血液を用いて痴呆症の検査を簡単に行うことができる方法を提供する。
【００１８】
【図面の簡単な説明】
【図１】 イムノクロマト法による本発明の実施の１例図である。
【図２】 ウエスタンブロット法によるアネキシンＶの検出結果である。
【符号の説明】
１、試料添加部
２、標識抗体保持部
３、展開部
４、抗アネキシンＶ抗体（抗アネキシンＶ抗体）固定部
５、抗アネキシンＩＶ抗体（抗アネキシンＶ抗体）抗体固定部
６、吸収パッド[0001]
BACKGROUND OF THE INVENTION
The present invention is used in the field of clinical examination, and in particular as a method for examining dementia.
[0002]
[Prior art]
Alzheimer's disease (AD) is primarily a cognitive dysfunction characterized by progressive neurodegenerative disease onset in old age. The degree of dementia in AD has been linked to the number of senile plaques (SP) found in the cerebral cortex of AD patients (eg, Non-Patent Document 1). And histologically, SP is surrounded by degenerative neurites and active astrocytes, and biochemically, SP contains a small insoluble fibrous protein called β-amyloid (eg, Non-patent document 2). The cause of AD is unknown, and there are currently few treatments for AD. Diagnosis of AD mainly includes exclusion diagnosis based on diagnostic criteria and brain image diagnosis, and there are few other objective methods . That is, there is no simple test method for differential AD diagnosis in patients before onset or after onset. Although
[0003]
[Patent Document 1]
JP-A-7-253429.
[Patent Document 2]
JP-A-7-72147.
[Non-Patent Document 1]
Nature, 1966, 209, 109.
[Non-Patent Document 2]
Biochem Biophys
Res Commun. 120, 885-890 (1984)
[Non-Patent Document 3]
Am J Pathol,
145, 640-649
(1994)
[Non-Patent Document 4]
Alcohol Clin
Exp Res, 25, 55S-58S (2001)
[0004]
[Problems to be solved by the invention]
An object of the present invention is to provide a method for testing dementia, which can be easily tested using body fluids such as blood that can be easily collected.
[0005]
[Means for determining the problem]
As a result of intensive studies on a simple test method for dementia, the present inventors can easily test for dementia because the concentration of annexins in blood, especially annexin V, is closely related to the onset of dementia. As a result , the present invention has been completed.
[0006]
The present invention has the following configuration.
1. A method for examining dementia, comprising measuring annexin V concentration in blood .
2. The method according to 1 above, wherein the blood is plasma .
3. A reagent or kit using the method according to 1-2 above.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to a test for dementia, and annexins in body fluids, especially
The present invention relates to a test method for dementia by measuring the concentration of annexin V. The present invention is carried out by immunologically measuring annexin V in blood, more preferably plasma, using specific antibodies (including whole antibodies or partial fragments having antibody activity). As the antibody to be used, either a polyclonal antibody or a monoclonal antibody can be used, and it is not particularly limited. As the immunological measurement method, known methods such as EIA, RIA, latex agglutination, immunoprecipitation, Western blot, dot blot, and immunochromatography are applied. Taking the EIA method for annexin V measurement as an example, annexin V in a test sample immobilized on a solid phase was reacted, and an enzyme-labeled anti-annexin V antibody was further reacted to be captured on the solid phase. By detecting the amount of labeled antibody, the concentration of annexin V in the sample can be measured.
[0008]
Further, using the biological properties of annexin V, for example, after capturing annexin V in a test sample on a solid phase on which phospholipid, preferably phosphatidylserine is immobilized, the labeled anti-annexin V antibody is reacted, It can also be measured by detecting the reaction amount.
[0009]
The present invention may be tested by combining the concentrations of Annexin V in the blood and other annexins such as Annexin IV. For example, calculating the ratio between the two, setting a threshold value for both measured values, and determining the diagnostic usefulness based on the determination result that both threshold values are exceeded, either one is exceeded, or neither is exceeded It is done by examining sex. Further, the measurement of annexin V and annexin IV is not necessarily the same test sample as long as the annexin V in the blood is measured . For example, a combination of different samples such as serum and plasma, cerebrospinal fluid and plasma, etc. Is possible.
[0010]
Annexin molecular species have been reported to change in expression in neuropsychiatric diseases such as dementia and alcoholism (for example, Non-
[0011]
The method of performing the measurement of annexin V and annexin IV in combination is of course the measurement of each concentration using an immunological technique constructed individually as described above, such as EIA, latex turbidimetry, etc. In addition to this, each antibody (including its fragment) is immobilized in one reaction vessel, and the test sample is allowed to react, and annexin V and annexin IV are contained in one reaction vessel. A method is also possible by capturing two kinds of molecules and detecting them with a labeled antibody in which an antibody against each molecule (including a fragment thereof) is labeled with a different labeling substance. Furthermore, it is also possible to detect two types of antigens simultaneously using an immunochromatography method (FIG. 1).
[0012]
【Example】
Examples of the present invention are shown below, but the present invention is not limited to the examples.
[Example 1]
(Construction of an annexin V concentration measurement ELISA)
A mouse monoclonal antibody was prepared using annexin V purified from human placenta as an immunogen. The prepared monoclonal antibody (clone No. 23) was immobilized on a 96-well ELISA microtiter plate to prepare an anti-annexin V antibody solid phase plate. A monoclonal antibody (clone No. 49) different from the clone used for the solid phase plate was labeled with biotin to prepare a labeled antibody. These were combined to construct an annexin V assay ELISA reagent.
[0013]
[Example 2]
(Examination by Western blot)
Annexin detection in plasma of 11 patients with dementia (Alzheimer's disease: 3 males, 6 females, cerebrovascular dementia: 2 females) and 10 controls (5 males, 5 females) by Western blot did. Each patient examined the intelligence score in advance on the Hasegawa simple intelligence evaluation scale. 1 mL of each plasma was diluted 10-fold with PBS buffer, 100 μg of liposome having a mixing ratio of phosphatidylserine and phosphatidylcholine of 1: 9 (molar ratio) was added, 6 mM calcium chloride was further added at a final concentration, and the mixture was incubated for 15 minutes. Thereafter, the precipitate fraction was collected by centrifugation, washed and then subjected to SDS-PAGE, transferred to a PVDF membrane , Western blotted with a rabbit-derived anti-annexin V antibody, and the relationship between the presence of annexin V in the blood and dementia was observed. Examined. None of the annexin V was detected in the control group, whereas in dementia, annexin V was detected in 10 of 11 cases (sensitivity 90.9%), and detection or measurement of annexin V in the blood was dementia It was found to be useful for the examination.
[0014]
[Example 3]
(Examination with enzyme immunoassay)
The annexin V concentration in the specimen examined in Example 2 was measured by the ELISA method prepared in Example 1. The results are shown in Table 1.
[0015]
[Table 1]
In the measurement by ELISA method, the annexin V concentration in the plasma of patients with dementia was significantly higher than that in the control group, and it was found that the measurement of the annexin V concentration in plasma was useful for testing dementia. .
[0017]
【The invention's effect】
The present invention provides a method capable of easily performing a test for dementia using blood.
[0018]
[Brief description of the drawings]
BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is an example of an embodiment of the present invention by immunochromatography.
FIG. 2 shows annexin V detection results by Western blotting.
[Explanation of symbols]
1,
Claims (2)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003043452A JP4264469B2 (en) | 2003-02-21 | 2003-02-21 | Testing method for dementia |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003043452A JP4264469B2 (en) | 2003-02-21 | 2003-02-21 | Testing method for dementia |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2004251794A JP2004251794A (en) | 2004-09-09 |
| JP4264469B2 true JP4264469B2 (en) | 2009-05-20 |
Family
ID=33026430
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2003043452A Expired - Fee Related JP4264469B2 (en) | 2003-02-21 | 2003-02-21 | Testing method for dementia |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4264469B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008020383A (en) * | 2006-07-14 | 2008-01-31 | Sapporo Medical Univ | Method for analyzing body fluid protein using liposome as ligand and method for preparing body fluid protein |
| JP5565546B2 (en) * | 2008-09-12 | 2014-08-06 | 北海道公立大学法人 札幌医科大学 | Temperature-responsive magnetic fine particles surface-modified with lipophilic molecules and compositions forming liposome-like structures containing the fine particles and amphiphilic molecules |
-
2003
- 2003-02-21 JP JP2003043452A patent/JP4264469B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JP2004251794A (en) | 2004-09-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US7101680B2 (en) | Method for detecting the presence of monomeric brain associated human glutamine synthetase | |
| JP6096813B2 (en) | Multi-biomarker set for breast cancer diagnosis, detection method thereof, and breast cancer diagnosis kit including antibody thereto | |
| JP7492711B2 (en) | Alzheimer's Disease Biomarkers | |
| JP2007517188A (en) | Immunosorbent blood test to assess paroxysmal brain firing | |
| Yu et al. | Distinction between bacterial and viral infections by serum measurement of human neutrophil lipocalin (HNL) and the impact of antibody selection | |
| JP4751555B2 (en) | Diagnostic assays for stroke | |
| WO1998040748A1 (en) | Diagnosing neurologic disorders | |
| AU2018247291A1 (en) | Biomarker-based methods and biochips for aiding the diagnosis of stroke | |
| JP2024019509A (en) | Diagnostic markers and diagnostic kits for Alzheimer's disease or presymptomatic Alzheimer's disease, methods for evaluating the amount of amyloid β protein accumulated in the brain, and in vitro methods for assisting in the detection of Alzheimer's disease or presymptomatic Alzheimer's disease in subjects | |
| WO2014185145A1 (en) | Biomarker for psychiatric and neurological disorders | |
| JP4264469B2 (en) | Testing method for dementia | |
| EP3872494A1 (en) | Blood biomarker for detecting deposition of amyloid beta in brains of groups with normal cognitive function and mild cognitive impairment | |
| US7070945B2 (en) | Process for determining the presence of monomeric brain associated human glutamine synthetase in patients exhibiting mild cognitive impairment | |
| JP3752219B2 (en) | Identification method of dementia disease | |
| RU2661022C2 (en) | Tropomyosin isoforms related to alzheimer's disease and moderate cognitive impairments | |
| JP2005513480A (en) | Diagnosis and treatment of dementia using thrombospondin | |
| CN118584103B (en) | CSF diagnostic markers and their application in central nervous system psychiatric lupus | |
| JP7569552B2 (en) | Biomarkers for dementia diagnosis | |
| CN117007819B (en) | Kit for assisting diagnosis of Alzheimer's disease | |
| KR102145438B1 (en) | A composition for predicting a risk of neurodegenerative diseases and a method for predicting neurodegenerative diseases using the same | |
| KR101250464B1 (en) | A method for the diagnosis of Alzheimer＇s disease from determination of PGCP concentration in human plasma | |
| JP7288428B2 (en) | Use of anti-CD14 antibody useful for presepsin measurement | |
| JP4388886B2 (en) | Method for detecting Alzheimer's disease | |
| AU2002252888A1 (en) | Process for differential diagnosis of alzheimer's dementia and device therefor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20051228 |
|
| A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20071031 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20071218 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20080215 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20080326 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20080520 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20080613 |
|
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20081028 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20081118 |
|
| R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120227 Year of fee payment: 3 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120227 Year of fee payment: 3 |
|
| S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313113 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120227 Year of fee payment: 3 |
|
| R360 | Written notification for declining of transfer of rights |
Free format text: JAPANESE INTERMEDIATE CODE: R360 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120227 Year of fee payment: 3 |
|
| R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120227 Year of fee payment: 3 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20150227 Year of fee payment: 6 |
|
| LAPS | Cancellation because of no payment of annual fees |