CN112114138A - 一种儿童肺炎支原体抗原的检测试剂盒及检测方法 - Google Patents
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Abstract
本发明公开了一种儿童肺炎支原体抗原的检测试剂盒及检测方法，本发明检测试剂盒应用酶联免疫分析法测定样本中钙卫蛋白的含量。将酶结合物温育后经洗涤去掉未结合物，彻底洗涤后加入底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。待测标本中钙卫蛋白浓度越高，标记抗原和抗体的结合就越受到抑制，显色愈浅。显色的深浅与酶量呈正相关，而与样本中待测钙卫蛋白含量呈负相关。用酶标仪在450nm波长下测定吸光度，计算样品浓度，进而判断样本中钙卫蛋白含量，为小儿肺炎支原体检测提供一种更直接、有效的检测方式，解决了现有技术中儿童肺炎支原体肺炎诊断方法比较单一的问题。
Description
技术领域
本发明涉及生物检测领域，具体涉及一种儿童肺炎支原体抗原的检测试剂盒及检测方法。
背景技术
肺炎支原体肺炎(MPP)又称原发性非典型肺炎、冷凝集阳性肺炎，为非典型肺炎中的一种，是由支原体(MP)感染引起的、呈间质性肺炎及毛细支气管炎样改变，占儿童社区获得性肺炎(CAP)的10％～40％。症状轻重不一。大多起病不急，有发热、厌食、咳嗽、畏寒、头痛、咽痛、胸骨下疼痛等症状，以发热和咳嗽为主要表现。体温在37℃～41℃，大多数在39℃左右，可为持续性或弛张性，或仅有低热，甚至不发热。多数咳嗽重，初期干咳，继而有痰(偶含少量血丝)，有时阵咳稍似百日咳。偶见恶心，呕吐及短暂的斑丘疹或荨麻疹。一般无呼吸困难表现，但婴儿患者可有喘鸣及呼吸困难。重症病例可合并胸腔积液、肺不张，或发生纵隔积气、气胸、坏死性肺炎等。少数病情发展迅速，可出现呼吸窘迫，甚至死亡。
诊断依据：
(1)临床表现。
(2)X线检查：肺部多种形态的浸润影，呈节段性分布，以肺下野多见，或从肺门附近向外伸展。病变常经3——4周后自行消散。
(3)实验室检查：①血白细胞总数正常或略增高，以中性粒为主；②冷凝集试验，起病2周后测定可阳性，若滴定效价大于1：32，尤其当滴度逐步升高时，更有诊断价值；③血清支原体IgM抗体的测定(酶联免疫吸附试验最敏感，免疫荧光法特异性强，间接血凝法较实用)阳性；④直接检测标本中肺炎支原体抗原，可用于临床早期快速诊断。所有患者的肺炎诊断均以临床和影像学检查为依据，肺炎支原体感染经血清学检查证实(IgM滴度≤1：160)。目前MPP缺乏直接、有效的预防和治疗策略，诊断方法比较单一。
发明内容
针对上述背景技术中不足，本发明的目的在于提供一种儿童肺炎支原体抗原的检测试剂盒，以解决现有技术中儿童肺炎支原体肺炎诊断方法比较单一的问题，为人肺炎支原体检测提供一种更直接、有效的检测方式。
为了实现以上目的，采用以下技术方案：
一种儿童肺炎支原体抗原的检测试剂盒，所述检测试剂盒中设有：预先包被已知钙卫蛋白抗体的ELISA板、酶结合物工作液、阳性质控、阴性质控、样品稀释液、生物素化抗体工作液、显色剂、浓缩清洗洗涤液和终止液；
其中所述的钙卫蛋白抗体为抗人S100钙结合蛋白A8、抗人S100钙结合蛋白P、抗人S100钙结合蛋白A9或抗人S100钙结合蛋白A12。
进一步地，所述阳性质控为肺炎支原体阳性血清，所述阴性质控为健康阴性血清。
进一步地，所述终止液为1mol/L的H2SO4溶液。
进一步地，所述浓缩清洗洗涤液为PBS-Tween缓冲液。
进一步地，所述显色剂为TMB。
本发明的另一个目的在于，提供一种使用上述的儿童肺炎支原体抗原的检测试剂盒的检测方法，其特征在于，包括以下步骤：
步骤a，采取待测样品以及阳性对照血清，阴性对照血清；
步骤b，加样：使用如权利要求1所述的检测试剂盒，分别设空白孔、标准孔、待测样品孔；空白孔加标准品&样品稀释液100μl，余孔分别加标准品或待测样品100μl；覆膜，于36±1℃孵育90分钟；
步骤c，弃去孔内液体，甩干，不用洗板，每孔中加入生物素化抗体工作液100μl(在使用前20分钟内配制)，覆膜，于36±1℃温育60分钟；
步骤d，弃去液体，洗板3次，每次浸泡30S，300-350μl/每孔，甩干并在吸水纸上轻拍将孔内液体拍干；
步骤e，每孔加酶结合物工作液(临用前20分钟内配制，避光放置)100μl，加上覆膜，36±1℃温育30分钟；弃去孔内液体，甩干，洗板5次；
步骤f，每孔加显色剂90μl，加上覆膜，36±1℃避光孵育15分钟；
步骤g，每孔加终止液50μl，终止反应，此时蓝色立转黄色；
步骤h，立即用酶标仪在450nm波长测量各孔的光密度。
与已有技术相比，本发明有益效果体现在：
本发明检测试剂盒应用酶联免疫分析法测定样本中钙卫蛋白的含量，通过检测盒检测钙卫蛋白浓度，进而判断肺炎支原体抗原。
将酶结合物温育后经洗涤去掉未结合物，彻底洗涤后加入底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。待测标本中钙卫蛋白浓度越高，标记抗原和抗体的结合就越受到抑制，显色愈浅。显色的深浅与酶量呈正相关，而与样本中待测钙卫蛋白含量呈负相关。用酶标仪在450nm波长下测定吸光度(OD值)，计算样品浓度，进而判断样本中钙卫蛋白含量，为小儿肺炎支原体检测提供一种更直接、有效的检测方式，解决了现有技术中儿童肺炎支原体肺炎诊断方法比较单一的问题。
具体实施方式
以下结合具体实施例对本发明的具体实施方式进行详细说明。应当理解的是，此处所描述的具体实施方式仅用于说明和解释本发明，并不用于限制本发明。
实施例1
一种儿童肺炎支原体抗原的检测试剂盒，所述检测试剂盒中设有：预先包被已知钙卫蛋白抗体的ELISA板、酶结合物工作液、阳性质控、阴性质控、样品稀释液、生物素化抗体工作液、显色剂、浓缩清洗洗涤液和终止液；
所述钙卫蛋白抗体——抗人S100钙结合蛋白A8、抗人S100钙结合蛋白P、抗人S100钙结合蛋白A9或抗人S100钙结合蛋白A12；
阳性质控——肺炎支原体阳性血清；阴性质控——健康阴性血清；
终止液——1mol/L的H2SO4溶液；
浓缩清洗洗涤液——PBS-Tween缓冲液；
显色剂——TMB。
实施例2
1、材料与方法
1.1研究对象
病例组：选择从2018年9月至2019年6月入住安徽医科大学第一附属医院儿科的患者100例，女42例，男58例，患者平均年龄为5.38岁，按儿童肺炎支原体肺炎的诊断标准进行划分。对照组：为同期门诊健康体检儿童，100例，女47例，男53例，平均年龄为5.21岁。
1.2方法
采集2ml静脉血，置于经抗凝剂处理过的真空采血管中，离心分离出血清，Elisa检测血清钙卫蛋白含量。
1.3统计分析
利用统计学SPSS软件进行数据处理与临床分析，计量资料以x±s表示，采用t检验，有统计学差异后分析与观察组临床指标间的关系，指标间的相关性分析采用线性回归；检验水准α＝0.05，以p﹤0.05为差异有统计学意义，探讨它们与疾病的之间的联系。
2、结果
根据统计结果可知，肺炎支原体阳性儿童血清S100A8/S100A9含量较健康儿童高，P<0.001。
**在0.01水平(双侧)上显著相关。
*在0.05水平(双侧)上显著相关。
分析发现，S100A8与体重比值和WBC、L、PLT正相关；
S100A8与体重比值和N、血红蛋白负相关；
S100A9与体重比值高同上。
3、讨论
肺炎支原体肺炎(MPP)又称原发性非典型肺炎、冷凝集阳性肺炎，为非典型肺炎中的一种，致病机制目前尚不明确。钙卫蛋白是钙离子结合蛋白，与钙离子结合，其理化性质极其稳定。在感染和炎症性疾病时，由于其特异性地在炎性细胞(中性粒细胞和巨噬细胞)中表达，因此，测定血液、分泌液和特定组织中钙卫蛋白的含量在一定程度上可以反映疾病的严重程度。通过肺炎支原体阳性儿童血清钙卫蛋白与健康儿童对比，发现钙卫蛋白与肺炎支原体肺炎具有相关性。对观察组临床数据分析发现，钙卫蛋白与患儿疾病相关临床指标具有相关性，可通过检测血清钙卫蛋白含量对肺炎支原体肺炎进行诊断，对疾病治疗进程和预后提供依据。
以上显示和描述了本发明的基本原理、主要特征和本发明的优点。本行业的技术人员应该了解，本发明不受上述实施例的限制，上述实施例和说明书中描述的只是本发明的原理，在不脱离本发明精神和范围的前提下本发明还会有各种变化和改进，这些变化和改进都落入要求保护的本发明的范围内。本发明要求的保护范围由所附的权利要求书及其等同物界定。
Claims (6)
1.一种儿童肺炎支原体抗原的检测试剂盒，其特征在于，所述检测试剂盒中设有：预先包被已知钙卫蛋白抗体的ELISA板、酶结合物工作液、阳性质控、阴性质控、样品稀释液、生物素化抗体工作液、显色剂、浓缩清洗洗涤液和终止液；
其中所述的钙卫蛋白抗体为抗人S100钙结合蛋白A8、抗人S100钙结合蛋白P、抗人S100钙结合蛋白A9或抗人S100钙结合蛋白A12。
2.根据权利要求1所述的一种儿童肺炎支原体抗原的检测试剂盒，其特征在于，所述阳性质控为肺炎支原体阳性血清，所述阴性质控为健康阴性血清。
3.根据权利要求1所述的一种儿童肺炎支原体抗原的检测试剂盒，其特征在于，所述终止液为1mol/L的H2SO4溶液。
4.根据权利要求1所述的一种儿童肺炎支原体抗原的检测试剂盒，其特征在于，所述浓缩清洗洗涤液为PBS-Tween缓冲液。
5.根据权利要求1所述的一种儿童肺炎支原体抗原的检测试剂盒，其特征在于，所述显色剂为TMB。
6.一种使用如权利要求1-5任一所述的儿童肺炎支原体抗原的检测试剂盒的检测方法，其特征在于，包括以下步骤：
步骤a，采取待测样品以及阳性对照血清，阴性对照血清；
步骤b，加样：使用如权利要求1所述的检测试剂盒，分别设空白孔、标准孔、待测样品孔；空白孔加标准品&样品稀释液100μl，余孔分别加标准品或待测样品100μl；覆膜，于36±1℃孵育90分钟；
步骤c，弃去孔内液体，甩干，不用洗板，每孔中加入生物素化抗体工作液100μl，覆膜，于36±1℃温育60分钟；
步骤d，弃去液体，洗板3次，每次浸泡30S，300-350μl/每孔，甩干并在吸水纸上轻拍将孔内液体拍干；
步骤e，每孔加酶结合物工作液100μl，加上覆膜，36±1℃温育30分钟；弃去孔内液体，甩干，洗板5次；
步骤f，每孔加显色剂90μl，加上覆膜，36±1℃避光孵育15分钟；
步骤g，每孔加终止液50μl，终止反应，此时蓝色立转黄色；
步骤h，立即用酶标仪在450nm波长测量各孔的光密度。
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013052844A1 (en) * | 2011-10-07 | 2013-04-11 | Pulmatrix, Inc. | Methods for treating and diagnosing respiratory tract infections |
| CN203479810U (zh) * | 2013-09-27 | 2014-03-12 | 武汉优尔生科技股份有限公司 | 小鼠s100钙结合蛋白b酶联免疫吸附测定试剂盒 |
| WO2019238531A1 (en) * | 2018-06-13 | 2019-12-19 | Westfälische Wilhelms-Universität Münster | Novel biomarkers for recurrent tonsillitis |
| US20200049706A1 (en) * | 2016-10-31 | 2020-02-13 | Peas Institut Ab | Device and method for detecting a bacterial infection |
| CN212321634U (zh) * | 2020-01-10 | 2021-01-08 | 陈红波 | 一种子痫前期相关蛋白psg9的检测试剂盒 |
-
2020
- 2020-07-29 CN CN202010743934.XA patent/CN112114138A/zh active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013052844A1 (en) * | 2011-10-07 | 2013-04-11 | Pulmatrix, Inc. | Methods for treating and diagnosing respiratory tract infections |
| CN203479810U (zh) * | 2013-09-27 | 2014-03-12 | 武汉优尔生科技股份有限公司 | 小鼠s100钙结合蛋白b酶联免疫吸附测定试剂盒 |
| US20200049706A1 (en) * | 2016-10-31 | 2020-02-13 | Peas Institut Ab | Device and method for detecting a bacterial infection |
| WO2019238531A1 (en) * | 2018-06-13 | 2019-12-19 | Westfälische Wilhelms-Universität Münster | Novel biomarkers for recurrent tonsillitis |
| CN212321634U (zh) * | 2020-01-10 | 2021-01-08 | 陈红波 | 一种子痫前期相关蛋白psg9的检测试剂盒 |
Non-Patent Citations (3)
| Title |
|---|
| HAVELKA, A 等: "Calprotectin, a new biomarker for diagnosis of acute respiratory infections", SCI REP, pages 4208 * |
| 丁圣刚;王亚亭;: "258例儿童支原体肺炎临床及并发症分析", 临床肺科杂志, no. 07, pages 741 - 742 * |
| 王凤霞 等: "钙卫蛋白的临床研究进展", 国际检验医学杂志, pages 971 - 973 * |
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