CN113777314A - 尿液踝蛋白1及其多肽片段在烧伤中的应用 - Google Patents

尿液踝蛋白1及其多肽片段在烧伤中的应用 Download PDF

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CN113777314A
CN113777314A CN202010519703.0A CN202010519703A CN113777314A CN 113777314 A CN113777314 A CN 113777314A CN 202010519703 A CN202010519703 A CN 202010519703A CN 113777314 A CN113777314 A CN 113777314A
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张曼
王佶图
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Abstract

本发明提供一种尿液踝蛋白1（Talin‑1）及其多肽片段的应用，具体为尿液踝蛋白1及其多肽片段在制备用于烧伤诊断、鉴别诊断、烧伤面积及程度评价、治疗效果评价、监测、预后评估及机理研究等制剂的应用。烧伤是日常生活中常见的重要创伤，每年每100万人中约有5000～100000人烧伤，据世界卫生组织统计，每年全球死于烧伤患者超过30万人，严重烧伤救治存活率仍处于较低水平。本发明通过研究证实，与健康人（正常对照组）相比，尿液踝蛋白1及其多肽片段在烧伤患者中表达升高。可用于烧伤患者的各种目的应用检测。本发明发挥尿液标本获取无创、可大规模重复取样、保存方便的优势，利用尿液标本检测尿液踝蛋白1及其多肽片段。

Description

尿液踝蛋白1及其多肽片段在烧伤中的应用
技术领域
本发明涉及尿液踝蛋白1及其多肽片段的新用途，具体涉及尿液踝蛋白1及其多肽片段在于烧伤诊断、鉴别诊断、烧伤面积及程度评价、治疗效果评价、监测、预后评估及机理研究等的应用。
背景技术
烧伤是指由于火焰、高温气体、灼热的固体或液体、放射线、电能、强酸强碱等化学物质所引起的人体皮肤、组织甚至深层脏器的损伤，是一种全身性综合性疾病。烧伤后创面组织大量坏死、感染、休克及凝血功能障碍等反应可引起机体发生一系列的病理生理学变化。不同程度的烧伤对人体有着不同的影响，严重的烧伤可对人体内环境造成破坏，使烧伤患者出现各系统复杂性的病理生理变化，相关的检测指标也会随着烧伤严重程度的不同随之出现相应的改变。及时检测这些指标的变化可为临床医生对于疾病的诊断，病情的判断，治疗方法的选择以及患者预后的评估等多方面提供有价值的参考依据。
但是，严重烧伤的病人皮肤完整性较差，血液学检测作为皮肤有创性检查在此类患者的临床应用中存在着诸多困难，反复的抽血检测还会加剧患者痛苦。尿液作为血液的超滤液蕴含着丰富的生物学信息，其采集过程具有无创便捷等优势，这在烧伤患者的检测中尤为明显。在尿液中寻找有助于烧伤诊断及反映病情变化的生物标志物，可提高烧伤患者的生活质量和依从性，减轻多次采血的痛苦，更好的为临床医生提供有利于疾病诊断和治疗的参考依据。
踝蛋白1（Talin-1）是一种大分子细胞骨架蛋白，由TLN1基因编码，在心肌细胞和骨骼肌细胞中广泛表达，在平滑肌和非肌肉的其他细胞的黏着斑中也存在着表达。目前研究表明，踝蛋白1一方面可介导整合素与细胞骨架肌动蛋白的偶联，另一方面还可与整合素结合促进黏着斑的形成，进而激活多条信号通路调控细胞的侵袭与转移。本研究烧伤患者尿液中踝蛋白1较健康人组出现表达上调，烧伤患者尿液中该蛋白含量增加。
与常用的临床血液样本相比，尿液可以完全无创、连续、大量收集；没有稳态调节，可累积更多种类、更大幅度的变化，机体的很多病理生理变化可能体现在尿液中。一些激素和细胞因子等分子量相对较小的蛋白多肽入血后，会很快被排泄进入尿液，这些蛋白和多肽在尿液中被检测到的概率比在血中大很多；尿液收集之前，尿中可能的蛋白降解过程已经完成，所以尿蛋白可在较长时间内保持稳定。为减轻烧伤患者多次采血的痛苦，本实验在前期方法学摸索的基础上，期望通过尿液蛋白或多肽研究，实现用无痛、方便、快捷、易重复的尿液检测辅助烧伤患者的诊断及病情监测，也为进一步尿液多肽检测试剂盒的研究奠定基础。
发明内容
本发明的目的在于提供一种尿液踝蛋白1及其多肽片段在制备用于烧伤诊断、鉴别诊断、烧伤面积及程度评价、治疗效果评价、监测、预后评估及机理研究等制剂的应用。
优选地，所述尿液踝蛋白1的氨基酸序列如SEQ ID NO.1所示(MVALSLKISIGNVVKTMQFE PSTMVYDACR IIRERIPEAP AGPPSDFGLF LSDDDPKKGI WLEAGKALDY YMLRNGDTMEYRKKQRPLKI RMLDGTVKTI MVDDSKTVTD MLMTICARIG ITNHDEYSLV RELMEEKKEE ITGTLRKDKTLLRDEKKMEK LKQKLHTDDE LNWLDHGRTL REQGVEEHET LLLRRKFFYS DQNVDSRDPV QLNLLYVQARDDILNGSHPV SFDKACEFAG FQCQIQFGPH NEQKHKAGFL DLKDFLPKEY VKQKGERKIF QAHKNCGQMSEIEAKVRYVK LARSLKTYGV SFFLVKEKMK GKNKLVPRLL GITKECVMRV DEKTKEVIQE WNLTNIKRWAASPKSFTLDF GDYQDGYYSV QTTEGEQIAQ LIAGYIDIIL KKKKSKDHFG LEGDEESTML EDSVSPKKSTVLQQQYNRVG KVEHGSVALP AIMRSGASGP ENFQVGSMPP AQQQITSGQM HRGHMPPLTS AQQALTGTINSSMQAVQAAQ ATLDDFDTLP PLGQDAASKA WRKNKMDESK HEIHSQVDAI TAGTASVVNL TAGDPAETDYTAVGCAVTTI SSNLTEMSRG VKLLAALLED EGGSGRPLLQ AAKGLAGAVS ELLRSAQPAS AEPRQNLLQAAGNVGQASGE LLQQIGESDT DPHFQDALMQ LAKAVASAAA ALVLKAKSVA QRTEDSGLQT QVIAAATQCALSTSQLVACT KVVAPTISSP VCQEQLVEAG RLVAKAVEGC VSASQAATED GQLLRGVGAA ATAVTQALNELLQHVKAHAT GAGPAGRYDQ ATDTILTVTE NIFSSMGDAG EMVRQARILA QATSDLVNAI KADAEGESDLENSRKLLSAA KILADATAKM VEAAKGAAAH PDSEEQQQRL REAAEGLRMA TNAAAQNAIK KKLVQRLEHAAKQAAASATQ TIAAAQHAAS TPKASAGPQP LLVQSCKAVA EQIPLLVQGV RGSQAQPDSP SAQLALIAASQSFLQPGGKM VAAAKASVPT IQDQASAMQL SQCAKNLGTA LAELRTAAQK AQEACGPLEM DSALSVVQNLEKDLQEVKAA ARDGKLKPLP GETMEKCTQD LGNSTKAVSS AIAQLLGEVA QGNENYAGIA ARDVAGGLRSLAQAARGVAA LTSDPAVQAI VLDTASDVLD KASSLIEEAK KAAGHPGDPE SQQRLAQVAK AVTQALNRCVSCLPGQRDVD NALRAVGDAS KRLLSDSLPP STGTFQEAQS RLNEAAAGLN QAATELVQAS RGTPQDLARASGRFGQDFST FLEAGVEMAG QAPSQEDRAQ VVSNLKGISM SSSKLLLAAK ALSTDPAAPN LKSQLAAAARAVTDSINQLI TMCTQQAPGQ KECDNALREL ETVRELLENP VQPINDMSYF GCLDSVMENS KVLGEAMTGISQNAKNGNLP EFGDAISTAS KALCGFTEAA AQAAYLVGVS DPNSQAGQQG LVEPTQFARA NQAIQMACQSLGEPGCTQAQ VLSAATIVAK HTSALCNSCR LASARTTNPT AKRQFVQSAK EVANSTANLV KTIKALDGAFTEENRAQCRA ATAPLLEAVD NLSAFASNPE FSSIPAQISP EGRAAMEPIV ISAKTMLESA GGLIQTARALAVNPRDPPSW SVLAGHSRTV SDSIKKLITS MRDKAPGQLE CETAIAALNS CLRDLDQASL AAVSQQLAPREGISQEALHT QMLTAVQEIS HLIEPLANAA RAEASQLGHK VSQMAQYFEP LTLAAVGAAS KTLSHPQQMALLDQTKTLAE SALQLLYTAK EAGGNPKQAA HTQEALEEAV QMMTEAVEDL TTTLNEAASA AGVVGGMVDSITQAINQLDE GPMGEPEGSF VDYQTTMVRT AKAIAVTVQE MVTKSNTSPE ELGPLANQLT SDYGRLASEAKPAAVAAENE EIGSHIKHRV QELGHGCAAL VTKAGALQCS PSDAYTKKEL IECARRVSEK VSHVLAALQAGNRGTQACIT AASAVSGIIA DLDTTIMFAT AGTLNREGTE TFADHREGIL KTAKVLVEDT KVLVQNAAGSQEKLAQAAQS SVATITRLAD VVKLGAASLG AEDPETQVVL INAVKDVAKA LGDLISATKA AAGKVGDDPAVWQLKNSAKV MVTNVTSLLK TVKAVEDEAT KGTRALEATT EHIRQELAVF CSPEPPAKTS TPEDFIRMTKGITMATAKAV AAGNSCRQED VIATANLSRR AIADMLRACK EAAYHPEVAP DVRLRALHYG RECANGYLELLDHVLLTLQK PSPELKQQLT GHSKRVAGSV TELIQAAEAM KGTEWVDPED PTVIAENELL GAAAAIEAAAKKLEQLKPRA KPKEADESLN FEEQILEAAK SIAAATSALV KAASAAQREL VAQGKVGAIP ANALDDGQWSQGLISAARMV AAATNNLCEA ANAAVQGHAS QEKLISSAKQ VAASTAQLLV ACKVKADQDS EAMKRLQAAGNAVKRASDNL VKAAQKAAAF EEQENETVVV KEKMVGGIAQ IIAAQEEMLR KERELEEARK KLAQIRQQQYKFLPSELRDE H）；或由SEQ ID NO.1所示的氨基酸序列衍生的，且与SEQ ID NO.1所示的氨基酸序列具有相同功能的氨基酸序列。
优选地，所述制剂为烧伤患者尿液踝蛋白1及其多肽片段检测试剂盒。
优选地，所述试剂盒包括抗原抗体反应的免疫方法及其试剂盒如能够特异性结合踝蛋白1及其多肽片段的适配体抗体或抗体片段中的一种或多种。
优选地，所述检测方法包括直接检测踝蛋白1及其多肽片段的质谱等方法及其相关试剂盒。
优选地，所述检测方法包括直接检测踝蛋白1及其多肽片段的相关核酸检测等方法及其相关试剂盒。
优选地，所述试剂盒还包括选自下组的成分：固相载体，稀释液，对照品，标准品，质控品，检测抗体，第二抗体、第二抗体稀释液，发光试剂，洗涤液、显色液、终止液中的任意一种或几种的组合。
优选地，所述标准品包括踝蛋白1标准品、人源化标签抗体标准品；较佳地，所述质控品包括：踝蛋白1质控品、人源化标签抗体质控品；较佳地，所述固相载体包括：微粒、微球、玻片、试纸条、塑料珠、液相芯片、微孔板或亲和膜等以及同等功能的其他载体。
优选地，所述固相载体的材质为聚氯乙烯、聚苯乙烯、聚丙酰胺、纤维素中的任意一种及具有类似功能的载体。
发明人首先收集了健康人、烧伤患者的尿液标本， 4000r/min离心5min后，吸取上清，采用Bradford法测定提取的蛋白浓度，进行SDS-PAGE酶解。尿液样本的Label-free质谱分析由OrbitrapFusion型质谱完成。将烧伤组和正常对照组在质谱中得到的数据进行定量计算。以蛋白表达量差异在1.5倍以上且经统计检验P<0.05作为参考标准筛选差异性多肽。然后发明人对具有统计学意义的差异性多肽进行鉴定，利用数据库检索得到差异蛋白踝蛋白1。
本发明通过研究证实与健康人相比，踝蛋白1及其多肽片段在烧伤患者的尿液中呈高表达，与临床诊断有较好的一致性。从而提出检测尿液踝蛋白1及其多肽片段可用于烧伤的辅助诊断或病情监测。
本发明发挥尿液标本获取无创、可大规模重复取样、保存方便的优势，利用尿液标本检测尿液踝蛋白1及其多肽片段。
为让本发明的上述和其它目的、特征和优点能更明显易懂，下文特举较佳实施例，并配合附图，作详细说明如下。
附图说明
图1是尿液踝蛋白1及其多肽片段在烧伤组及健康对照组中含量图。
图2是踝蛋白1参与主要生物过程示意图。
具体实施方式
实施例1 尿液标本的收集与处理
选取烧伤患者作为烧伤组，选取同期健康体检者作为正常对照组。收集各组研究对象入院后的新鲜晨尿样本30ml，不能正常排尿者收集其早晨导尿管中尿液，置于干燥、洁净的容器内。将收集的尿液标本4000r/min离心5min后，吸取上清，每管2ml分装，-80℃冰箱保存。
实施例2 质谱分析和尿液多肽的筛选
对尿液样品蛋白提取，并对提取的蛋白浓度进行测定。尿液样本的质谱分析由OrbitrapFusion型质谱完成。将实验组和正常对照组在质谱中得到的数据进行定量计算。组间比较采用t-test进行差异分析，以蛋白表达量差异在1.5倍以上且经统计检验P<0.05作为参考标准筛选差异表达蛋白。
实施例3 差异多肽的鉴定及分析
使用的数据库为Uniprot_Homo数据库，产生的质谱原始文件采用MaxQuant软件处理，检索参数设置见表1。
Figure 516288DEST_PATH_IMAGE001
与健康人相比，踝蛋白1在烧伤患者的尿液中高表达，如图1所示，其参与的主要生物过程如图2所示，踝蛋白1在正常对照组和烧伤组尿液中的表达有显著性差异。
虽然本发明已以较佳实施例披露如上，然其并非用以限定本发明，任何所属技术领域的技术人员，在不脱离本发明的精神和范围内，当可作些许的更动与改进，因此本发明的保护范围当视权利要求所界定者为准。
序列表
<110> 张, 曼
<120> 尿液踝蛋白1及其多肽片段在烧伤中的应用
<130> 1
<140> 20PTalin-1
<141> 2020-05-10
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2541
<212> PRT
<213> Human Urine
<400> 1
Met Val Ala Leu Ser Leu Lys Ile Ser Ile Gly Asn Val Val Lys Thr
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Met Gln Phe Glu Pro Ser Thr Met Val Tyr Asp Ala Cys Arg Ile Ile
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Arg Glu Arg Ile Pro Glu Ala Pro Ala Gly Pro Pro Ser Asp Phe Gly
35 40 45
Leu Phe Leu Ser Asp Asp Asp Pro Lys Lys Gly Ile Trp Leu Glu Ala
50 55 60
Gly Lys Ala Leu Asp Tyr Tyr Met Leu Arg Asn Gly Asp Thr Met Glu
65 70 75 80
Tyr Arg Lys Lys Gln Arg Pro Leu Lys Ile Arg Met Leu Asp Gly Thr
85 90 95
Val Lys Thr Ile Met Val Asp Asp Ser Lys Thr Val Thr Asp Met Leu
100 105 110
Met Thr Ile Cys Ala Arg Ile Gly Ile Thr Asn His Asp Glu Tyr Ser
115 120 125
Leu Val Arg Glu Leu Met Glu Glu Lys Lys Glu Glu Ile Thr Gly Thr
130 135 140
Leu Arg Lys Asp Lys Thr Leu Leu Arg Asp Glu Lys Lys Met Glu Lys
145 150 155 160
Leu Lys Gln Lys Leu His Thr Asp Asp Glu Leu Asn Trp Leu Asp His
165 170 175
Gly Arg Thr Leu Arg Glu Gln Gly Val Glu Glu His Glu Thr Leu Leu
180 185 190
Leu Arg Arg Lys Phe Phe Tyr Ser Asp Gln Asn Val Asp Ser Arg Asp
195 200 205
Pro Val Gln Leu Asn Leu Leu Tyr Val Gln Ala Arg Asp Asp Ile Leu
210 215 220
Asn Gly Ser His Pro Val Ser Phe Asp Lys Ala Cys Glu Phe Ala Gly
225 230 235 240
Phe Gln Cys Gln Ile Gln Phe Gly Pro His Asn Glu Gln Lys His Lys
245 250 255
Ala Gly Phe Leu Asp Leu Lys Asp Phe Leu Pro Lys Glu Tyr Val Lys
260 265 270
Gln Lys Gly Glu Arg Lys Ile Phe Gln Ala His Lys Asn Cys Gly Gln
275 280 285
Met Ser Glu Ile Glu Ala Lys Val Arg Tyr Val Lys Leu Ala Arg Ser
290 295 300
Leu Lys Thr Tyr Gly Val Ser Phe Phe Leu Val Lys Glu Lys Met Lys
305 310 315 320
Gly Lys Asn Lys Leu Val Pro Arg Leu Leu Gly Ile Thr Lys Glu Cys
325 330 335
Val Met Arg Val Asp Glu Lys Thr Lys Glu Val Ile Gln Glu Trp Asn
340 345 350
Leu Thr Asn Ile Lys Arg Trp Ala Ala Ser Pro Lys Ser Phe Thr Leu
355 360 365
Asp Phe Gly Asp Tyr Gln Asp Gly Tyr Tyr Ser Val Gln Thr Thr Glu
370 375 380
Gly Glu Gln Ile Ala Gln Leu Ile Ala Gly Tyr Ile Asp Ile Ile Leu
385 390 395 400
Lys Lys Lys Lys Ser Lys Asp His Phe Gly Leu Glu Gly Asp Glu Glu
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Ser Thr Met Leu Glu Asp Ser Val Ser Pro Lys Lys Ser Thr Val Leu
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Gln Gln Gln Tyr Asn Arg Val Gly Lys Val Glu His Gly Ser Val Ala
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Leu Pro Ala Ile Met Arg Ser Gly Ala Ser Gly Pro Glu Asn Phe Gln
450 455 460
Val Gly Ser Met Pro Pro Ala Gln Gln Gln Ile Thr Ser Gly Gln Met
465 470 475 480
His Arg Gly His Met Pro Pro Leu Thr Ser Ala Gln Gln Ala Leu Thr
485 490 495
Gly Thr Ile Asn Ser Ser Met Gln Ala Val Gln Ala Ala Gln Ala Thr
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Leu Asp Asp Phe Asp Thr Leu Pro Pro Leu Gly Gln Asp Ala Ala Ser
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Lys Ala Trp Arg Lys Asn Lys Met Asp Glu Ser Lys His Glu Ile His
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Ser Gln Val Asp Ala Ile Thr Ala Gly Thr Ala Ser Val Val Asn Leu
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Thr Ala Gly Asp Pro Ala Glu Thr Asp Tyr Thr Ala Val Gly Cys Ala
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Val Thr Thr Ile Ser Ser Asn Leu Thr Glu Met Ser Arg Gly Val Lys
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Leu Leu Ala Ala Leu Leu Glu Asp Glu Gly Gly Ser Gly Arg Pro Leu
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Leu Gln Ala Ala Lys Gly Leu Ala Gly Ala Val Ser Glu Leu Leu Arg
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Ser Ala Gln Pro Ala Ser Ala Glu Pro Arg Gln Asn Leu Leu Gln Ala
625 630 635 640
Ala Gly Asn Val Gly Gln Ala Ser Gly Glu Leu Leu Gln Gln Ile Gly
645 650 655
Glu Ser Asp Thr Asp Pro His Phe Gln Asp Ala Leu Met Gln Leu Ala
660 665 670
Lys Ala Val Ala Ser Ala Ala Ala Ala Leu Val Leu Lys Ala Lys Ser
675 680 685
Val Ala Gln Arg Thr Glu Asp Ser Gly Leu Gln Thr Gln Val Ile Ala
690 695 700
Ala Ala Thr Gln Cys Ala Leu Ser Thr Ser Gln Leu Val Ala Cys Thr
705 710 715 720
Lys Val Val Ala Pro Thr Ile Ser Ser Pro Val Cys Gln Glu Gln Leu
725 730 735
Val Glu Ala Gly Arg Leu Val Ala Lys Ala Val Glu Gly Cys Val Ser
740 745 750
Ala Ser Gln Ala Ala Thr Glu Asp Gly Gln Leu Leu Arg Gly Val Gly
755 760 765
Ala Ala Ala Thr Ala Val Thr Gln Ala Leu Asn Glu Leu Leu Gln His
770 775 780
Val Lys Ala His Ala Thr Gly Ala Gly Pro Ala Gly Arg Tyr Asp Gln
785 790 795 800
Ala Thr Asp Thr Ile Leu Thr Val Thr Glu Asn Ile Phe Ser Ser Met
805 810 815
Gly Asp Ala Gly Glu Met Val Arg Gln Ala Arg Ile Leu Ala Gln Ala
820 825 830
Thr Ser Asp Leu Val Asn Ala Ile Lys Ala Asp Ala Glu Gly Glu Ser
835 840 845
Asp Leu Glu Asn Ser Arg Lys Leu Leu Ser Ala Ala Lys Ile Leu Ala
850 855 860
Asp Ala Thr Ala Lys Met Val Glu Ala Ala Lys Gly Ala Ala Ala His
865 870 875 880
Pro Asp Ser Glu Glu Gln Gln Gln Arg Leu Arg Glu Ala Ala Glu Gly
885 890 895
Leu Arg Met Ala Thr Asn Ala Ala Ala Gln Asn Ala Ile Lys Lys Lys
900 905 910
Leu Val Gln Arg Leu Glu His Ala Ala Lys Gln Ala Ala Ala Ser Ala
915 920 925
Thr Gln Thr Ile Ala Ala Ala Gln His Ala Ala Ser Thr Pro Lys Ala
930 935 940
Ser Ala Gly Pro Gln Pro Leu Leu Val Gln Ser Cys Lys Ala Val Ala
945 950 955 960
Glu Gln Ile Pro Leu Leu Val Gln Gly Val Arg Gly Ser Gln Ala Gln
965 970 975
Pro Asp Ser Pro Ser Ala Gln Leu Ala Leu Ile Ala Ala Ser Gln Ser
980 985 990
Phe Leu Gln Pro Gly Gly Lys Met Val Ala Ala Ala Lys Ala Ser Val
995 1000 1005
Pro Thr Ile Gln Asp Gln Ala Ser Ala Met Gln Leu Ser Gln Cys Ala
1010 1015 1020
Lys Asn Leu Gly Thr Ala Leu Ala Glu Leu Arg Thr Ala Ala Gln Lys
1025 1030 1035 1040
Ala Gln Glu Ala Cys Gly Pro Leu Glu Met Asp Ser Ala Leu Ser Val
1045 1050 1055
Val Gln Asn Leu Glu Lys Asp Leu Gln Glu Val Lys Ala Ala Ala Arg
1060 1065 1070
Asp Gly Lys Leu Lys Pro Leu Pro Gly Glu Thr Met Glu Lys Cys Thr
1075 1080 1085
Gln Asp Leu Gly Asn Ser Thr Lys Ala Val Ser Ser Ala Ile Ala Gln
1090 1095 1100
Leu Leu Gly Glu Val Ala Gln Gly Asn Glu Asn Tyr Ala Gly Ile Ala
1105 1110 1115 1120
Ala Arg Asp Val Ala Gly Gly Leu Arg Ser Leu Ala Gln Ala Ala Arg
1125 1130 1135
Gly Val Ala Ala Leu Thr Ser Asp Pro Ala Val Gln Ala Ile Val Leu
1140 1145 1150
Asp Thr Ala Ser Asp Val Leu Asp Lys Ala Ser Ser Leu Ile Glu Glu
1155 1160 1165
Ala Lys Lys Ala Ala Gly His Pro Gly Asp Pro Glu Ser Gln Gln Arg
1170 1175 1180
Leu Ala Gln Val Ala Lys Ala Val Thr Gln Ala Leu Asn Arg Cys Val
1185 1190 1195 1200
Ser Cys Leu Pro Gly Gln Arg Asp Val Asp Asn Ala Leu Arg Ala Val
1205 1210 1215
Gly Asp Ala Ser Lys Arg Leu Leu Ser Asp Ser Leu Pro Pro Ser Thr
1220 1225 1230
Gly Thr Phe Gln Glu Ala Gln Ser Arg Leu Asn Glu Ala Ala Ala Gly
1235 1240 1245
Leu Asn Gln Ala Ala Thr Glu Leu Val Gln Ala Ser Arg Gly Thr Pro
1250 1255 1260
Gln Asp Leu Ala Arg Ala Ser Gly Arg Phe Gly Gln Asp Phe Ser Thr
1265 1270 1275 1280
Phe Leu Glu Ala Gly Val Glu Met Ala Gly Gln Ala Pro Ser Gln Glu
1285 1290 1295
Asp Arg Ala Gln Val Val Ser Asn Leu Lys Gly Ile Ser Met Ser Ser
1300 1305 1310
Ser Lys Leu Leu Leu Ala Ala Lys Ala Leu Ser Thr Asp Pro Ala Ala
1315 1320 1325
Pro Asn Leu Lys Ser Gln Leu Ala Ala Ala Ala Arg Ala Val Thr Asp
1330 1335 1340
Ser Ile Asn Gln Leu Ile Thr Met Cys Thr Gln Gln Ala Pro Gly Gln
1345 1350 1355 1360
Lys Glu Cys Asp Asn Ala Leu Arg Glu Leu Glu Thr Val Arg Glu Leu
1365 1370 1375
Leu Glu Asn Pro Val Gln Pro Ile Asn Asp Met Ser Tyr Phe Gly Cys
1380 1385 1390
Leu Asp Ser Val Met Glu Asn Ser Lys Val Leu Gly Glu Ala Met Thr
1395 1400 1405
Gly Ile Ser Gln Asn Ala Lys Asn Gly Asn Leu Pro Glu Phe Gly Asp
1410 1415 1420
Ala Ile Ser Thr Ala Ser Lys Ala Leu Cys Gly Phe Thr Glu Ala Ala
1425 1430 1435 1440
Ala Gln Ala Ala Tyr Leu Val Gly Val Ser Asp Pro Asn Ser Gln Ala
1445 1450 1455
Gly Gln Gln Gly Leu Val Glu Pro Thr Gln Phe Ala Arg Ala Asn Gln
1460 1465 1470
Ala Ile Gln Met Ala Cys Gln Ser Leu Gly Glu Pro Gly Cys Thr Gln
1475 1480 1485
Ala Gln Val Leu Ser Ala Ala Thr Ile Val Ala Lys His Thr Ser Ala
1490 1495 1500
Leu Cys Asn Ser Cys Arg Leu Ala Ser Ala Arg Thr Thr Asn Pro Thr
1505 1510 1515 1520
Ala Lys Arg Gln Phe Val Gln Ser Ala Lys Glu Val Ala Asn Ser Thr
1525 1530 1535
Ala Asn Leu Val Lys Thr Ile Lys Ala Leu Asp Gly Ala Phe Thr Glu
1540 1545 1550
Glu Asn Arg Ala Gln Cys Arg Ala Ala Thr Ala Pro Leu Leu Glu Ala
1555 1560 1565
Val Asp Asn Leu Ser Ala Phe Ala Ser Asn Pro Glu Phe Ser Ser Ile
1570 1575 1580
Pro Ala Gln Ile Ser Pro Glu Gly Arg Ala Ala Met Glu Pro Ile Val
1585 1590 1595 1600
Ile Ser Ala Lys Thr Met Leu Glu Ser Ala Gly Gly Leu Ile Gln Thr
1605 1610 1615
Ala Arg Ala Leu Ala Val Asn Pro Arg Asp Pro Pro Ser Trp Ser Val
1620 1625 1630
Leu Ala Gly His Ser Arg Thr Val Ser Asp Ser Ile Lys Lys Leu Ile
1635 1640 1645
Thr Ser Met Arg Asp Lys Ala Pro Gly Gln Leu Glu Cys Glu Thr Ala
1650 1655 1660
Ile Ala Ala Leu Asn Ser Cys Leu Arg Asp Leu Asp Gln Ala Ser Leu
1665 1670 1675 1680
Ala Ala Val Ser Gln Gln Leu Ala Pro Arg Glu Gly Ile Ser Gln Glu
1685 1690 1695
Ala Leu His Thr Gln Met Leu Thr Ala Val Gln Glu Ile Ser His Leu
1700 1705 1710
Ile Glu Pro Leu Ala Asn Ala Ala Arg Ala Glu Ala Ser Gln Leu Gly
1715 1720 1725
His Lys Val Ser Gln Met Ala Gln Tyr Phe Glu Pro Leu Thr Leu Ala
1730 1735 1740
Ala Val Gly Ala Ala Ser Lys Thr Leu Ser His Pro Gln Gln Met Ala
1745 1750 1755 1760
Leu Leu Asp Gln Thr Lys Thr Leu Ala Glu Ser Ala Leu Gln Leu Leu
1765 1770 1775
Tyr Thr Ala Lys Glu Ala Gly Gly Asn Pro Lys Gln Ala Ala His Thr
1780 1785 1790
Gln Glu Ala Leu Glu Glu Ala Val Gln Met Met Thr Glu Ala Val Glu
1795 1800 1805
Asp Leu Thr Thr Thr Leu Asn Glu Ala Ala Ser Ala Ala Gly Val Val
1810 1815 1820
Gly Gly Met Val Asp Ser Ile Thr Gln Ala Ile Asn Gln Leu Asp Glu
1825 1830 1835 1840
Gly Pro Met Gly Glu Pro Glu Gly Ser Phe Val Asp Tyr Gln Thr Thr
1845 1850 1855
Met Val Arg Thr Ala Lys Ala Ile Ala Val Thr Val Gln Glu Met Val
1860 1865 1870
Thr Lys Ser Asn Thr Ser Pro Glu Glu Leu Gly Pro Leu Ala Asn Gln
1875 1880 1885
Leu Thr Ser Asp Tyr Gly Arg Leu Ala Ser Glu Ala Lys Pro Ala Ala
1890 1895 1900
Val Ala Ala Glu Asn Glu Glu Ile Gly Ser His Ile Lys His Arg Val
1905 1910 1915 1920
Gln Glu Leu Gly His Gly Cys Ala Ala Leu Val Thr Lys Ala Gly Ala
1925 1930 1935
Leu Gln Cys Ser Pro Ser Asp Ala Tyr Thr Lys Lys Glu Leu Ile Glu
1940 1945 1950
Cys Ala Arg Arg Val Ser Glu Lys Val Ser His Val Leu Ala Ala Leu
1955 1960 1965
Gln Ala Gly Asn Arg Gly Thr Gln Ala Cys Ile Thr Ala Ala Ser Ala
1970 1975 1980
Val Ser Gly Ile Ile Ala Asp Leu Asp Thr Thr Ile Met Phe Ala Thr
1985 1990 1995 2000
Ala Gly Thr Leu Asn Arg Glu Gly Thr Glu Thr Phe Ala Asp His Arg
2005 2010 2015
Glu Gly Ile Leu Lys Thr Ala Lys Val Leu Val Glu Asp Thr Lys Val
2020 2025 2030
Leu Val Gln Asn Ala Ala Gly Ser Gln Glu Lys Leu Ala Gln Ala Ala
2035 2040 2045
Gln Ser Ser Val Ala Thr Ile Thr Arg Leu Ala Asp Val Val Lys Leu
2050 2055 2060
Gly Ala Ala Ser Leu Gly Ala Glu Asp Pro Glu Thr Gln Val Val Leu
2065 2070 2075 2080
Ile Asn Ala Val Lys Asp Val Ala Lys Ala Leu Gly Asp Leu Ile Ser
2085 2090 2095
Ala Thr Lys Ala Ala Ala Gly Lys Val Gly Asp Asp Pro Ala Val Trp
2100 2105 2110
Gln Leu Lys Asn Ser Ala Lys Val Met Val Thr Asn Val Thr Ser Leu
2115 2120 2125
Leu Lys Thr Val Lys Ala Val Glu Asp Glu Ala Thr Lys Gly Thr Arg
2130 2135 2140
Ala Leu Glu Ala Thr Thr Glu His Ile Arg Gln Glu Leu Ala Val Phe
2145 2150 2155 2160
Cys Ser Pro Glu Pro Pro Ala Lys Thr Ser Thr Pro Glu Asp Phe Ile
2165 2170 2175
Arg Met Thr Lys Gly Ile Thr Met Ala Thr Ala Lys Ala Val Ala Ala
2180 2185 2190
Gly Asn Ser Cys Arg Gln Glu Asp Val Ile Ala Thr Ala Asn Leu Ser
2195 2200 2205
Arg Arg Ala Ile Ala Asp Met Leu Arg Ala Cys Lys Glu Ala Ala Tyr
2210 2215 2220
His Pro Glu Val Ala Pro Asp Val Arg Leu Arg Ala Leu His Tyr Gly
2225 2230 2235 2240
Arg Glu Cys Ala Asn Gly Tyr Leu Glu Leu Leu Asp His Val Leu Leu
2245 2250 2255
Thr Leu Gln Lys Pro Ser Pro Glu Leu Lys Gln Gln Leu Thr Gly His
2260 2265 2270
Ser Lys Arg Val Ala Gly Ser Val Thr Glu Leu Ile Gln Ala Ala Glu
2275 2280 2285
Ala Met Lys Gly Thr Glu Trp Val Asp Pro Glu Asp Pro Thr Val Ile
2290 2295 2300
Ala Glu Asn Glu Leu Leu Gly Ala Ala Ala Ala Ile Glu Ala Ala Ala
2305 2310 2315 2320
Lys Lys Leu Glu Gln Leu Lys Pro Arg Ala Lys Pro Lys Glu Ala Asp
2325 2330 2335
Glu Ser Leu Asn Phe Glu Glu Gln Ile Leu Glu Ala Ala Lys Ser Ile
2340 2345 2350
Ala Ala Ala Thr Ser Ala Leu Val Lys Ala Ala Ser Ala Ala Gln Arg
2355 2360 2365
Glu Leu Val Ala Gln Gly Lys Val Gly Ala Ile Pro Ala Asn Ala Leu
2370 2375 2380
Asp Asp Gly Gln Trp Ser Gln Gly Leu Ile Ser Ala Ala Arg Met Val
2385 2390 2395 2400
Ala Ala Ala Thr Asn Asn Leu Cys Glu Ala Ala Asn Ala Ala Val Gln
2405 2410 2415
Gly His Ala Ser Gln Glu Lys Leu Ile Ser Ser Ala Lys Gln Val Ala
2420 2425 2430
Ala Ser Thr Ala Gln Leu Leu Val Ala Cys Lys Val Lys Ala Asp Gln
2435 2440 2445
Asp Ser Glu Ala Met Lys Arg Leu Gln Ala Ala Gly Asn Ala Val Lys
2450 2455 2460
Arg Ala Ser Asp Asn Leu Val Lys Ala Ala Gln Lys Ala Ala Ala Phe
2465 2470 2475 2480
Glu Glu Gln Glu Asn Glu Thr Val Val Val Lys Glu Lys Met Val Gly
2485 2490 2495
Gly Ile Ala Gln Ile Ile Ala Ala Gln Glu Glu Met Leu Arg Lys Glu
2500 2505 2510
Arg Glu Leu Glu Glu Ala Arg Lys Lys Leu Ala Gln Ile Arg Gln Gln
2515 2520 2525
Gln Tyr Lys Phe Leu Pro Ser Glu Leu Arg Asp Glu His
2530 2535 2540

Claims (9)

1.尿液踝蛋白1及其多肽片段在制备用于烧伤诊断、鉴别诊断、烧伤面积及程度评价、治疗效果评价、监测、预后评估及机理研究等制剂的应用。
2.根据权利要求1所述的应用，其特征在于，所述尿液踝蛋白1的氨基酸序列如SEQ IDNO.1所示；或由SEQ ID NO.1所示的氨基酸序列衍生的，且与SEQ ID NO.1所示的氨基酸序列具有相同功能的氨基酸序列。
3.根据权利要求1所述的应用，其特征在于，所述制剂为烧伤患者尿液踝蛋白1及其多肽片段检测试剂盒。
4.根据权利要求3所述的应用，其特征在于，所述试剂盒包括抗原抗体反应的免疫方法及其试剂盒如能够特异性结合踝蛋白1及其多肽片段的适配体抗体或抗体片段中的一种或多种。
5.根据权利要求3所述的应用，其特征在于，所述检测方法包括直接检测踝蛋白1及其多肽片段的质谱等方法及其相关试剂盒。
6.根据权利要求3所述的应用，其特征在于，所述检测方法包括直接检测踝蛋白1及其多肽片段或其相关核酸检测等方法及其相关试剂盒。
7.根据权利要求3所述的应用，其特征在于，所述试剂盒还包括选自下组的成分：固相载体，稀释液，对照品，标准品，质控品，检测抗体，第二抗体、第二抗体稀释液，发光试剂，洗涤液、显色液、终止液中的任意一种或几种的组合。
8.根据权利要求7所述的应用，其特征在于，所述标准品包括踝蛋白1标准品、人源化标签抗体标准品；较佳地，所述质控品包括：踝蛋白1控品、人源化标签抗体质控品；较佳地，所述固相载体包括：微粒、微球、玻片、试纸条、塑料珠、液相芯片、微孔板或亲和膜等以及同等功能的其他载体。
9.根据权利要求8所述的应用，其特征在于，所述固相载体的材质为聚氯乙烯、聚苯乙烯、聚丙酰胺、纤维素中的任意一种及具有类似功能的载体。
CN202010519703.0A 2020-06-09 2020-06-09 尿液踝蛋白1及其多肽片段在烧伤中的应用 Pending CN113777314A (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114895023A (zh) * 2022-05-07 2022-08-12 浙江大学 检测抗Talin-1-IgG自身抗体的试剂在制备检测血管内皮损伤的试剂盒中的应用

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114895023A (zh) * 2022-05-07 2022-08-12 浙江大学 检测抗Talin-1-IgG自身抗体的试剂在制备检测血管内皮损伤的试剂盒中的应用

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