WO2021213322A1 - 一种检测肾癌患者外周血循环肿瘤细胞pd-l1表达的免疫荧光试剂盒及检测方法 - Google Patents
一种检测肾癌患者外周血循环肿瘤细胞pd-l1表达的免疫荧光试剂盒及检测方法 Download PDFInfo
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Abstract
Description
Claims (6)
- 一种检测肾癌患者外周血循环肿瘤细胞PD-L1基因突变的试剂盒,其特征在于,包括稀释液45mL,脱色液1mL,染色液A 0.5mL,染色液B 1mL,200μl甲醇、200μl 2%PFA,100μl 10%山羊血清,小鼠抗CK、大鼠抗CD45和兔抗PD-L1组成的一抗混悬液100μl,荧光标记的羊抗小鼠、荧光标记的羊抗大鼠、荧光标记的羊抗兔组成的二抗混悬液100μL,DAPI封片剂;一抗混悬液中小鼠抗CK、大鼠抗CD45和兔抗PD-L1分别按1:100、1:400和1:500稀释,总体积为100μL;二抗混悬液中荧光标记的羊抗小鼠、荧光标记的羊抗大鼠、荧光标记的羊抗兔按1:500稀释。
- 根据权利要求1所述的试剂盒,其特征在于,所述稀释液是由1mmol/L EDTA+0.1%BSA+0.1%海藻糖+0.2%月桂醇聚氧乙烯醚组成。
- 根据权利要求1所述的试剂盒,其特征在于,所述脱色液是由95%酒精与100%二甲苯按容积比1:1组成。
- 根据权利要求1所述的试剂盒,其特征在于,所述染色液A为DAB染色液;所述染色液B为苏木素染色液。
- 一种利用权利要求1-4任一项所述的试剂盒非诊断目的检测肾癌患者外周血循环肿瘤细胞PD-L1表达的方法,其特征在于,包括以下步骤:(1)利用膜过滤装置分离获取无法获得组织标本的晚期或复发肾癌患者外周血:采集无法获取组织标本的晚期或复发肾癌患者肘正中静脉外周血5ml;(2)外周血样预处理:将采集的外周血样采用稀释液进行10倍稀释,稀释后加多聚甲醛固定外周血样10分钟,固定终浓度为0.25%;(3)利用膜过滤分离肿瘤细胞装置过滤外周血样,分离获得外周血CTC:将预处理的外周血样加入到膜过滤分离肿瘤细胞装置的血样容器中,使其依靠重力自然过滤;(4)过滤结束后,从膜过滤分离肿瘤细胞装置中取下滤器,将循环肿瘤细胞染色液A液0.5ml加入到滤器中,染色3min,PBS缓冲液冲洗干净;滤液过滤完全后加入染色液B液1ml,染色2min,纯水1ml冲洗2次;(5)向滤器中加入200μl 2%PFA,室温固定5min,完成后0.5ml PBS漂洗3次,每次2min;(6)再向滤器中加入200μl预冷的甲醇,4℃固定15min,取下滤膜,放置在载玻片上,干燥后在显微镜下观察,确定是否存在CTC;(7)运用免疫荧光方法检测外周血CTC的PD-L1表达情况。
- 根据权利要求5所述的检测方法,其特征在于,步骤(7)所述检测外周血CTC的PD-L1表达的具体方法如下:(1)脱色:将带有CTC的滤膜从载玻片上取下,置于脱色液中浸泡4-6小时,脱去CTC染色液,PBS洗2min×3次;(2)封闭:向滤膜上滴加100μl 10%山羊血清,室温放置30min,完成后吸去多余的血清(注:羊血清用PBS稀释);(3)一抗孵育:向滤膜上滴加100μl小鼠抗CK、大鼠抗CD45和兔抗PD-L1组成的一抗混悬液,37℃孵育1h或4℃过夜,完成后PBS洗3min×3次;(4)二抗孵育:向滤膜上滴加100μl荧光标记的羊抗小鼠、荧光标记的羊抗大鼠、荧光标记的羊抗兔组成的二抗混悬液,室温孵育30min,完成后PBS洗2min×3次;(5)使用含DAPI的封片剂封片,阅片,采图;(6)采图完成后,脱片后进行瑞氏吉姆萨染色,与IF结果进行对比。
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Citations (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0123443A2 (en) * | 1983-03-24 | 1984-10-31 | Fuji Photo Film Co., Ltd. | Method of analyzing whole blood sample |
WO2010005991A2 (en) * | 2008-07-07 | 2010-01-14 | The Board Of Regents Of The University Of Texas System | Circulating tumor and tumor stem cell detection using genomic specific probes |
CN105209919A (zh) * | 2013-03-15 | 2015-12-30 | 豪夫迈·罗氏有限公司 | 治疗pd-1和pd-l1相关疾患的生物标志物和方法 |
WO2016061064A1 (en) * | 2014-10-15 | 2016-04-21 | Epic Sciences, Inc. | Circulating tumor cell diagnostics for therapy targeting pd-l1 |
CN105548569A (zh) * | 2016-01-28 | 2016-05-04 | 山东省医学科学院 | 一种肾癌患者外周血vegf的检测方法 |
WO2017072539A1 (en) * | 2015-10-27 | 2017-05-04 | Pharmassist Ltd | Method for the quantification of pd-l1 expression |
US20180095086A1 (en) * | 2016-10-03 | 2018-04-05 | Hitachi Chemical Company, Ltd. | Standard cell suspension |
CN108507992A (zh) * | 2018-04-09 | 2018-09-07 | 苏州大学附属第医院 | 循环肿瘤细胞表面标志分子pd-l1的检测方法 |
CN111141906A (zh) * | 2020-01-06 | 2020-05-12 | 中南大学湘雅医院 | 一种小细胞肺癌患者外周血循环肿瘤细胞及pd-l1的检测试剂盒 |
CN111521792A (zh) * | 2020-04-21 | 2020-08-11 | 山东第一医科大学(山东省医学科学院) | 检测非小细胞肺癌患者外周血循环肿瘤细胞pd-l1基因突变的免疫荧光试剂盒及方法 |
CN111521796A (zh) * | 2020-04-21 | 2020-08-11 | 山东第一医科大学(山东省医学科学院) | 一种检测肾癌患者外周血循环肿瘤细胞pd-l1表达的免疫荧光试剂盒及检测方法 |
CN111551718A (zh) * | 2020-04-20 | 2020-08-18 | 山东第一医科大学(山东省医学科学院) | 一种检测前列腺癌患者外周血循环肿瘤细胞pd-l1表达的免疫荧光试剂盒及检测方法 |
CN111638359A (zh) * | 2020-07-01 | 2020-09-08 | 山东凯歌智能机器有限公司 | 检测小细胞肺癌患者外周血循环肿瘤细胞pd-l1基因突变的免疫荧光试剂盒及检测方法 |
CN111679077A (zh) * | 2020-07-01 | 2020-09-18 | 山东凯歌智能机器有限公司 | 肾细胞癌患者外周血循环肿瘤细胞E-Cadherin表达的免疫荧光试剂盒及检测方法 |
-
2020
- 2020-04-21 CN CN202010316999.6A patent/CN111521796A/zh not_active Withdrawn
-
2021
- 2021-04-19 WO PCT/CN2021/088116 patent/WO2021213322A1/zh active Application Filing
Patent Citations (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0123443A2 (en) * | 1983-03-24 | 1984-10-31 | Fuji Photo Film Co., Ltd. | Method of analyzing whole blood sample |
WO2010005991A2 (en) * | 2008-07-07 | 2010-01-14 | The Board Of Regents Of The University Of Texas System | Circulating tumor and tumor stem cell detection using genomic specific probes |
CN105209919A (zh) * | 2013-03-15 | 2015-12-30 | 豪夫迈·罗氏有限公司 | 治疗pd-1和pd-l1相关疾患的生物标志物和方法 |
WO2016061064A1 (en) * | 2014-10-15 | 2016-04-21 | Epic Sciences, Inc. | Circulating tumor cell diagnostics for therapy targeting pd-l1 |
WO2017072539A1 (en) * | 2015-10-27 | 2017-05-04 | Pharmassist Ltd | Method for the quantification of pd-l1 expression |
CN105548569A (zh) * | 2016-01-28 | 2016-05-04 | 山东省医学科学院 | 一种肾癌患者外周血vegf的检测方法 |
US20180095086A1 (en) * | 2016-10-03 | 2018-04-05 | Hitachi Chemical Company, Ltd. | Standard cell suspension |
CN108507992A (zh) * | 2018-04-09 | 2018-09-07 | 苏州大学附属第医院 | 循环肿瘤细胞表面标志分子pd-l1的检测方法 |
CN111141906A (zh) * | 2020-01-06 | 2020-05-12 | 中南大学湘雅医院 | 一种小细胞肺癌患者外周血循环肿瘤细胞及pd-l1的检测试剂盒 |
CN111551718A (zh) * | 2020-04-20 | 2020-08-18 | 山东第一医科大学(山东省医学科学院) | 一种检测前列腺癌患者外周血循环肿瘤细胞pd-l1表达的免疫荧光试剂盒及检测方法 |
CN111521792A (zh) * | 2020-04-21 | 2020-08-11 | 山东第一医科大学(山东省医学科学院) | 检测非小细胞肺癌患者外周血循环肿瘤细胞pd-l1基因突变的免疫荧光试剂盒及方法 |
CN111521796A (zh) * | 2020-04-21 | 2020-08-11 | 山东第一医科大学(山东省医学科学院) | 一种检测肾癌患者外周血循环肿瘤细胞pd-l1表达的免疫荧光试剂盒及检测方法 |
CN111638359A (zh) * | 2020-07-01 | 2020-09-08 | 山东凯歌智能机器有限公司 | 检测小细胞肺癌患者外周血循环肿瘤细胞pd-l1基因突变的免疫荧光试剂盒及检测方法 |
CN111679077A (zh) * | 2020-07-01 | 2020-09-18 | 山东凯歌智能机器有限公司 | 肾细胞癌患者外周血循环肿瘤细胞E-Cadherin表达的免疫荧光试剂盒及检测方法 |
Non-Patent Citations (2)
Title |
---|
GALATEA KALLERGI, ELENI-KYRIAKI VETSIKA, DESPOINA AGGOURAKI, ELENI LAGOUDAKI, ANASTASIOS KOUTSOPOULOS, FILIPPOS KOINIS, PANAGIOTIS: "Evaluation of PD-L1/PD-1 on circulating tumor cells in patients with advanced non-small cell lung cancer", THERAPEUTIC ADVANCES IN MEDICAL ONCOLOGY, vol. 10, 1 January 2018 (2018-01-01), pages 1 - 11, XP055859904, DOI: 10.1177/1758834017750121 * |
HAN LU;LI SHENG;LU ZUO-WEI;LIU MIN;LI HAO;LU XIN-YANG: "Morphology of Circulating Tumor Cells in Gastric Cancer and Esophageal Cancer", CHINESE JOURNAL OF DIAGNOSTIC PATHOLOGY, vol. 24, no. 10, 28 October 2017 (2017-10-28), pages 778 - 782, XP055859893, ISSN: 1007-8096, DOI: 10.3969/j.issn.1007-8096.2017.10.014 * |
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