CN105548546B - 一种肺癌筛查试剂盒 - Google Patents
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Abstract
本发明公开了一种肺癌筛查试剂盒，它包括任选的用于检测BAI‑1表达水平的试剂。本发明还公开了检测BAI‑1表达水平的试剂在制备肺癌筛查用试剂中的用途。本发明试剂盒通过检测BAI‑1的表达水平，可以判断待检人群患肺癌的风险，可用于临床肺癌的辅助诊断，为患者采取相关的治疗措施或者决策提供有效的依据，临床应用前景良好。
Description
技术领域
本发明涉及一种肺癌筛查试剂盒。
背景技术
肺癌是世界上最常见的恶性肿瘤之一，其发病率和死亡率呈逐年上升趋势，目前发病率居世界首位，严重威胁着人类健康和生命。
肺癌的病因复杂，一般认为影响因素包括：①吸烟；②环境污染：如雾霾、室内装修；③不良生活方式：如饮食习惯差、生活压力大；④慢性肺部疾病：如肺结核、尘肺，矽肺、慢性支气管炎；⑤人体内在因素：如家族遗传、免疫机能降低、内分泌功能失调等。
同时，肺癌是一种善于隐匿的疾病，经常在疾病发展到晚期才表现出临床症状，70～80％的肺癌患者在诊断出患有肺癌症状时已是中、晚期，癌细胞已经扩散，错过了最佳治愈时机，五年生存率低。对于早期的肺癌患者，经过及时治疗可大大提高患者的5年及以上生存率和生存质量。因此肺癌的早期诊断和进行有效的筛查至关重要。
肺癌的筛查，是指对那些没有肺癌相关症状的人群进行常规体检，在出现症状前及时发现肺癌。如果可以找到血液里面的肺癌分子标志物，用于提示临床医生早期对患者采取相关的治疗措施或者决策具有重要的意义。
脑特异性新生血管抑制因子1(BAI-1，Brain angiogenesis inhibitor 1)，又称脑血管生成抑制剂1或脑血管生成抑制因子1，其作用是抑制血管生成。目前未见关于脑特异性新生血管抑制因子1与肺癌相关的报道。
发明内容
为了解决上述问题，发明人对肺癌进行了详细研究，发现了BAI-1可以作为其分子标志物。其中，BAI-1在血清中的表达水平与肺癌呈正相关。因此，通过检测血清中BAI-1的表达水平，可以预测待检人群患肺癌的风险。
据此，本发明提供了一种肺癌筛查试剂盒，以及检测BAI-1的表达水平的试剂在制备肺癌筛查试剂中的用途。
本发明的肺癌筛查试剂盒，它包括任选的用于检测BAI-1表达水平的试剂。
其中，所述试剂是用于检测血清中BAI-1表达水平的试剂。
其中，所述检测BAI-1表达水平的试剂为ELISA检测方法用试剂。
其中，所述检测BAI-1表达水平的试剂为Western Blot检测方法用试剂。
本发明还提供了检测BAI-1表达水平的试剂在制备肺癌筛查用试剂中的用途。
其中，所述试剂是用于检测血清中BAI-1表达水平的试剂。
其中，所述检测BAI-1表达水平的试剂为ELISA检测方法用试剂。
其中，所述检测BAI-1表达水平的试剂为Western Blot检测方法用试剂。
本发明试剂盒通过检测BAI-1的表达水平，可以判断待检人群患肺癌的风险：若BAI-1的表达水平高，则患肺癌的风险高，若BAI-1的表达水平低，则患肺癌的风险低，可用于临床肺癌的辅助诊断，临床应用前景良好。
显然，根据本发明的上述内容，按照本领域的普通技术知识和惯用手段，在不脱离本发明上述基本技术思想前提下，还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式，对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1为肺癌患者与健康对照血清检测结果图
图2为BAI-1敏感度及特异性分析图
具体实施方式
实施例1 BAI-1表达水平与肺癌的关系
一、实验方法
1、临床资料
选取肺癌患者60例，健康对照34例，基本信息如下：
*注：其他包括大细胞癌、腺鳞癌、小细胞癌。
2、BAI-1表达水平的检测(定量检测BAI-1的含量)
抽取肺癌患者及健康对照的血液，EDTA抗凝后离心(1200rpm，10min)，将上清吸取后进行第二次离心(3000rpm，15min)，吸取第二次离心后的上清，作为检测标本，分装后-80℃保存。
采用在BIOMATIK公司购买的BAI-1ELISA试剂盒(货号：EKU02798)，按照如下方法检测BAI-1的表达水平：
(1)标准品、检测标本100μl/孔加到抗体包被板中，用样品稀释液为空白对照，封板后37℃孵育2小时；
(2)弃上清液后加入100μl/孔检测液体A(检测液体A配制如下：将检测试剂A与检测试剂A稀释液按1:100比例混合)，封板后37℃孵育1小时；
(3)弃上清液，加入350μl洗涤液(Wash buffer)洗涤3次，吸水纸上将残余液体拍干；
(4)加入100μl/孔检测液体B(检测液体B配制如下：将检测试剂B与检测试剂B稀释液按1:100比例混合)，封板后37℃孵育30分钟；
(5)重复步骤(3)；
(6)加入90μl/孔TMB底物反应液，封板后37℃避光孵育15-25分钟；
(7)加入50μl/孔终止液后在酶标仪上450nm波长读板。
3、结果分析
(1)样本浓度：酶标仪根据标准品浓度及吸光值，自动算出各样品浓度；
(2)差异性分析：采用SPSS17.0对实验组(肺癌患者)和对照组(健康体检人群)进行统计分析；
(3)ROC分析：采用SPSS17.0对实验组(肺癌患者)和对照组(健康体检人群)进行分析；
(4)图片制作：用GraphPad Prism 5.0软件完成。
4、BAI-1的表达水平与肺癌的相关性
肺癌患者与健康对照血清中BAI-1的表达水平检测结果见图1。
由图1可见，肺癌患者的血清BAI-1平均表达水平为1926.3pg/ml，健康对照的血清BAI-1平均表达水平为689.3pg/ml，BAI-1在肺癌患者中显著升高，与健康对照组相比，BAI-1表达水平差异具有统计学意义(P<0.0001)。
BAI-1敏感度及特异性分析见图2。
由图2可见，BAI-1的最佳阈值(cut-off值)为>952pg/ml，曲线下面积(AUC)为92.6％，与健康对照组相比，BAI-1诊断特异性为85.3％，敏感度为95.0％。特异性和敏感度较高。
由以上结果可以看出，与正常人群相比，肺癌患者的BAI-1表达水平显著升高(P<0.0001)，说明肺癌与BAI-1表达水平呈正相关，BAI-1的高表达会显著提高患肺癌的可能性，因此，可以通过检测待检人群的BAI-1的表达水平，将肺癌的易感人群筛查出来。
实施例2本发明检测血液BAI-1表达水平的试剂盒的组成及其使用方法
一、ELISA检测试剂盒
1、试剂盒的组成
检测试剂盒(50人份)：
| 组分 | 体积 |
| BAI-1抗体包被板 | 1个 |
| BAI-1标准品 | 1ml(20000pg/ml) |
| 检测试剂A | 120ul |
| 检测试剂B | 120ul |
| 检测试剂A稀释液 | 12ml |
| 检测试剂B稀释液 | 12ml |
| TMB底物反应液 | 9ml |
| 洗涤液 | 20ml |
| 终止液 | 6ml |
2、试剂盒的使用方法
抽取待检样本血液，EDTA抗凝后离心(1200rpm，10min)，将上清吸取后进行第二次离心(3000rpm，15min)，吸取第二次离心后的上清，作为检测标本，按照如下方法检测BAI-1的表达水平：
(1)标准品、检测标本100μl/孔加到抗体包被板中，用样品稀释液为空白对照，封板后37℃孵育2小时；
(2)弃上清液后加入100μl/孔检测液体A(检测液体A配制如下：将检测试剂A与检测试剂A稀释液按1:100比例混合)，封板后37℃孵育1小时；
(3)弃上清液，加入350μl洗涤液(Wash buffer)洗涤3次，吸水纸上将残余液体拍干；
(4)加入100μl/孔检测液体B(检测液体B配制如下：将检测试剂B与检测试剂B稀释液按1:100比例混合)，封板后37℃孵育30分钟；
(5)重复步骤(3)；
(6)加入90μl/孔TMB底物反应液，封板后37℃避光孵育15-25分钟；
(7)加入50μl/孔终止液后在酶标仪上450nm波长读板，根据标准品浓度及吸光值，自动算出各样品浓度。
综上，本发明试剂盒通过检测BAI-1的表达水平，可以筛查待检人群患肺癌的风险：若BAI-1的表达水平高，则患肺癌的风险高，若BAI-1的表达水平低，则患肺癌的风险低，可用于临床肺癌的辅助诊断，为患者采取相关的治疗措施或者决策提供有效的依据，临床应用前景良好。
Claims (4)
1.检测脑特异性新生血管抑制因子1表达水平的试剂在制备肺癌筛查用试剂中的用途。
2.根据权利要求1所述的用途，其特征在于：所述试剂是用于检测血清中脑特异性新生血管抑制因子1表达水平的试剂。
3.根据权利要求1或2所述的用途，其特征在于：所述检测脑特异性新生血管抑制因子1表达水平的试剂为ELISA检测方法用试剂。
4.根据权利要求1或2所述的用途，其特征在于：所述检测脑特异性新生血管抑制因子1表达水平的试剂为Western Blot检测方法用试剂。
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| CN110873798A (zh) * | 2019-12-09 | 2020-03-10 | 四川大学华西医院 | Ppfia4自身抗体检测试剂在制备肺癌筛查试剂盒中的用途 |
| CN112415198B (zh) * | 2020-11-20 | 2022-11-11 | 四川大学华西医院 | Gp1bb检测试剂在制备肺癌筛查试剂盒中的用途 |
| CN115993453A (zh) * | 2022-07-22 | 2023-04-21 | 四川大学 | 用于rdaa阳性疾病的诊断和治疗的方法和试剂盒 |
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| WO2012088290A2 (en) * | 2010-12-22 | 2012-06-28 | Abbott Laboratories | Tri-variable domain binding proteins and uses thereof |
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