CN105548566A - 一种快速检测结核分枝杆菌的方法 - Google Patents

一种快速检测结核分枝杆菌的方法 Download PDF

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CN105548566A
CN105548566A CN201511028937.0A CN201511028937A CN105548566A CN 105548566 A CN105548566 A CN 105548566A CN 201511028937 A CN201511028937 A CN 201511028937A CN 105548566 A CN105548566 A CN 105548566A
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mycobacterium tuberculosis
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苏锐
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HEFEI ANWEIKANG MED LAB Co Ltd
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G01N33/6866—Interferon
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56911—Bacteria
    • G01N33/5695—Mycobacteria
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/555—Interferons [IFN]
    • G01N2333/57—IFN-gamma

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Abstract

本发明提供一种快速检测结核分枝杆菌的方法，涉及医学检测技术领域，步骤为：向受检血样中加入结核分枝杆菌特有的T细胞抗原刺激，于37℃环境下培养16～24小时后检测T细胞产生的γ干扰素的情况，如果T细胞由于结核分枝杆菌特异抗原的刺激而提高了γ干扰素的产量，并到达一定的阀值，则认为受检血样感染了结核分枝杆菌。本发明所用刺激抗原为结核分枝杆菌特有抗原，特异性高，不受BCG接种与非结核分枝杆菌的干扰；灵敏度好，非人体试验，结果判定客观，且不会对机体免疫状态产生影响。

Description

一种快速检测结核分枝杆菌的方法
技术领域
本发明涉及医学检测技术领域，尤其涉及一种快速检测结核分枝杆菌的方法。
背景技术
目前结核分枝杆菌的检测主要依据结核菌素进行检测(PPD检测)，然而PPD检测无法区分BCG接种产生的免疫应答反应与致病性结核分枝杆菌感染产生的应答反应，这让核分枝杆菌的检测有了难度，检测敏感度和特异性较低。
发明内容
本发明的目的在于提供一种快速检测结核分枝杆菌的方法，以解决上述技术问题。
本发明所要解决的技术问题采用以下技术方案来实现：
一种快速检测结核分枝杆菌的方法，其特征在于，步骤为：
向受检血样中加入结核分枝杆菌特有的T细胞抗原刺激，于37℃环境下培养16～24小时后检测T细胞产生的γ干扰素的情况，如果T细胞由于结核分枝杆菌特异抗原的刺激而提高了γ干扰素的产量，并到达一定的阀值，则认为受检血样感染了结核分枝杆菌。
结核分枝杆菌感染机体后，机体可产生具有抗原特异性的致敏T细胞，当机体再次受到结核分枝杆菌抗原刺激后，致敏T细胞可快速活化与增殖，释放出γ-干扰素。
是以RD1编码的结核特异性抗原为刺激原检测机体内致敏T淋巴细胞释放出干扰素。与PPD试验用的结核菌素不同，这些蛋白在所有结核分枝杆菌及致病性牛分枝杆菌中特异表达，而在卡介苗和大多数非结核分枝杆菌中不表达。因此，本发明检测结核分枝杆菌具有较高的敏感度和特异性。
γ-干扰素(IFN-γ)体外释放对于结核分枝杆菌感染，特别是区别自然感染和卡介苗接种等潜伏感染的检测具有较强的特异性。γ-干扰素体外释放通过特异性抗原刺激刺激血液淋巴细胞检测细胞因子，检测结核分枝杆菌的特异度和敏感度达到90％以上。
本发明的有益效果是：
第一，本发明所用刺激抗原为结核分枝杆菌特有抗原，而非结核分枝杆菌提取物，特异性高，不受BCG接种与非结核分枝杆菌的干扰，同时也不受导致类结核病症状的其他致病菌的影响。
第二，本发明检测结核分枝杆菌的方法灵敏度好，可应用于体外检测结核病，不仅对细菌学检查阳性结核病患者检出率较高，对菌阴及其他结核病患者也有90％以上的检出率，而菌阴及其他结核病患者恰恰是目前临床诊断的难点。
第三，本发明为非人体试验，结果判定客观，且不会对机体免疫状态产生影响。
本发明检测IFN-γ浓度的高低与结核分枝杆菌的多少及活性相关。
具体实施方式
为了使本发明实现的技术手段、创作特征、达成目的与功效易于明白了解，下面结合具体实施例，进一步阐述本发明，但下述实施例仅仅为本发明的优选实施例，并非全部。基于实施方式中的实施例，本领域技术人员在没有做出创造性劳动的前提下所获得其它实施例，都属于本发明的保护范围。
一种快速检测结核分枝杆菌的方法，
向受检血样中加入结核分枝杆菌特有的T细胞抗原刺激，于37℃环境下培养16～24小时后检测T细胞产生的γ干扰素的情况，如果T细胞由于结核分枝杆菌特异抗原的刺激而提高了γ干扰素的产量，并到达一定的阀值，则认为受检血样感染了结核分枝杆菌。
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仅为本发明的优选例，并不用来限制本发明，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范围内。本发明要求保护范围由所附的权利要求书及其等效物界定。

Claims (1)

1.一种快速检测结核分枝杆菌的方法，其特征在于，步骤为：
向受检血样中加入结核分枝杆菌特有的T细胞抗原刺激，于37℃环境下培养16～24小时后检测T细胞产生的γ干扰素的情况，如果T细胞由于结核分枝杆菌特异抗原的刺激而提高了γ干扰素的产量，并到达一定的阀值，则认为受检血样感染了结核分枝杆菌。
CN201511028937.0A 2015-12-25 2015-12-25 一种快速检测结核分枝杆菌的方法 Pending CN105548566A (zh)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101493454A (zh) * 2008-01-21 2009-07-29 范雄林 结核病抗原特异的全血IFN-γ诊断试剂盒及其制作方法和应用方法
CN102033129A (zh) * 2009-09-29 2011-04-27 上海英伯肯医学生物技术有限公司 用抗原刺激的细胞免疫反应来检测病原微生物的方法及测试笔
CN104569390A (zh) * 2015-01-26 2015-04-29 河北省胸科医院 一种γ-干扰素定量检测方法及试剂盒

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101493454A (zh) * 2008-01-21 2009-07-29 范雄林 结核病抗原特异的全血IFN-γ诊断试剂盒及其制作方法和应用方法
CN102033129A (zh) * 2009-09-29 2011-04-27 上海英伯肯医学生物技术有限公司 用抗原刺激的细胞免疫反应来检测病原微生物的方法及测试笔
CN104569390A (zh) * 2015-01-26 2015-04-29 河北省胸科医院 一种γ-干扰素定量检测方法及试剂盒

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