JP2018109646A - 脳梗塞の診断マーカー - Google Patents
脳梗塞の診断マーカー Download PDFInfo
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Abstract
Description
配列番号113 SOSTDC1-156 (biotin-KITVVTACKCKRYTR)
配列番号79 CTNND1-211 (biotin-LRNVSSERSEARRKL)
配列番号57 CLDND1-69 (biotin-FRYNGTVGLWRRCIT)
配列番号63 CCNG2-231 (biotin-KKHSKINDTEFFYWR)
即ち、FOXJ2(配列番号171)、SOSTDC1(配列番号113)、CTNND1(配列番号79)、CLDND1(配列番号57)、CCNG2(配列番号63)は脳梗塞によく反応するペプチドである。
エンザイムイムノアッセイ法は、被験試料中の合成ペプチド抗原に対する自己抗体を、固相に固定化された抗原タンパク質と反応させ、ヒト免疫グロブリンに対する標識二次抗体を添加し、自己抗体に結合した標識二次抗体を測定することによって行う。
凝集反応法は、被験試料中の脳梗塞又は心筋梗塞特異抗原に対する自己抗体を、担体粒子上に固定化された本発明の抗原ペプチドと反応させ、該抗原ペプチドと自己抗体との反応により生じる凝集を測定することにより行う。担体粒子としては、例えばポリスチレンラテックス、カオリン、ベントナイト、炭素末、ヒツジ、ニワトリ等の赤血球等を使用することができる。抗原ペプチドを担体粒子上に固定化する方法は公知の方法を適用することができる。例えば、タンニン酸、グルタルアルデヒド、ビスアゾベンジジン、カルボジイミド類、キノン類、塩化クロム類等のカップリング剤を使用する方法、物理的吸着による方法等により行うことができる。
較することにより被験試料中の自己抗体を定量することができる。
イムノクロマト法は毛細管現象を応用した免疫測定法であり、例えばサンドイッチ法を利用したイムノクロマト法では、対象抗原に特異的に結合する第1抗体を特定の領域に固定した不溶性薄膜状支持体(例えば、ガラス繊維膜、ナイロン膜、又はセルロ−ス膜など)中に、対象抗原と特異的に結合する標識化第2抗体と、対象抗原を含む可能性のある検体溶液とを展開し、不溶性薄膜状
支持体の第1抗体を固定した領域上で、対象抗原との免疫複合体を形成させ、標識の着色又は発色等の信号を検出し、対象抗原を測定する。なお、標識としては、例えば、酵素を含むタンパク質、着色ラテックス粒子、金属コロイド、又は炭素粒子を使用することができる。
い。
(1)プロトアレイを用いた一次スクリーニング
SLE患者血清は本人又は家族の同意(インフォームドコンセント)を得て、独立行政法人 国立病院機構 下志津病院において採取した。脳梗塞患者血清、心筋梗塞患者血清、糖尿病患者血清及び健常者検体は千葉大学附属病院及び下志津病院において同様に採取した。
配列番号79 CTNND1-211 (biotin-LRNVSSERSEARRKL)
配列番号57 CLDND1-69 (biotin-FRYNGTVGLWRRCIT)
配列番号63 CCNG2-231 (biotin-KKHSKINDTEFFYWR)
配列番号171 FOXJ2-426 (biotin-KMVNRLNWSSIEQSQ)
配列番号55 TFAM-231 (biotin-LRRTIKKQRKYGAEE)
配列番号128 TOP3B-628 (biotin-HRFMKYIQAKPSRLH)
配列番号87 MYBBP1A-1134 (biotin-LYWQAMKTLGVQRPK)
配列番号88 MYBBP1A-1306 (biotin-IRSPSLLQSGAKKKA)
配列番号76 KIF12-203 (biotin-CVSPSAQCLPETLST)
結果を表6及び表7に示す。表6及び表7は、AlphaLISA法で調べた血清抗体レベルを示すものであり、SLE患者と健常者とを比較して示している。表では、各グループの平均値、SD、平均値+2SD(カットオフ値)、総数、陽性数、陽性率が示されている。健常者検体の値の平均値+2SDをカットオフ値に設定した時の各グループの陽性数(P Positive No.)と陽性率(P Positive (%))も示されている。P(SLE vs HD)は、健常者検体と各グループの t testのP値を示す。
次に、上記で選別されたペプチド抗原について、高純度(90%以上)のペプチドを合成した。それらに対する脳梗塞又は心筋梗塞の患者血清における抗体レベルを AlphaLISA 法により測定した。
Claims (2)
- 配列番号171に示すアミノ酸配列からなるペプチドである脳梗塞の診断マーカー。
- 配列番号171で表わされるアミノ酸配列において、1若しくは数個のアミノ酸が欠失、置換若しくは付加されたアミノ酸配列からなるペプチドである請求項1に記載の脳梗塞の診断マーカー。
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