量子点荧光微球免疫层析试纸条定量检测恶性疟原虫

量子点荧光微球免疫层析试纸条定量检测恶性疟原虫

Quantum-dot Submicrobead-Based Immunochromatographic Strip for Rapid and Quantitative Detection of Plasmodium Falciparum

摘要:

以羧基化CdTe/ZnSe量子点荧光微球为荧光标记物,采用1-乙基-(3-二甲基氨基丙基)碳酰二亚胺(EDC)法偶联抗恶性疟原虫富组氨酸蛋白(Pf)单克隆抗体制备荧光探针;以羊抗恶性疟原虫组氨酸多克隆抗体和驴抗鼠二抗分别喷涂硝酸纤维膜,形成试纸条检测线和质控线,建立了免疫层析试纸条定量检测血清中恶性疟原虫的方法。所使用的羧基化量子点荧光微球的荧光强度为单个量子点的2800倍。实验结果表明,该荧光试纸条定量检测血清中恶性疟原虫线性范围为5.8~8010 Parasite/μL,最低灵敏度达到5.8 Parasite/μL,单个样品检测时间只需15 min。加标回收实验显示,试纸条批内回收率为93.0%~111.8%,批间回收率为98.3%~115.1%,且批内、批间的相对标准偏差均小于5%。

Abstract:

A CdTe/ZnSe quantum-dot submicrobead ( QBs ) , which exhibited fluorescence intensity approximately 2800-fold stronger than that of single quantum dots, was conjugated with the anti-histidine rich protein( HRP )-Ⅱ mAbs using N-( 3-( Dimethylamino ) propyl )-N’-ethylcarbodiimide hydrochloride ( EDC ) method as fluorescence probe. The goat anti-HRP-Ⅱ polyclonal antibodies and donkey anti-mouse polyclonal antibodies were sprayed onto the nitrocellulose membrane as test line and control line, respectively. The resultant fluorescence probes were introduced to the immunochromatographic strip for the quantitative determination of Plasmodium falciparum. For determination of Plasmodium falciparum in serum, the QBs based immunochromatographic strips exhibited a good dynamic linear range from 5 . 8 Parasite/μL to 8010 Parasite/μL with a limit of detection of 5. 8 Parasite/μL. The detection time of the proposed QBs based immunochromatographic strips for each sample was only 15 min. Moreover, the recovery rates of the intra-and inter-assay ranged from 93. 0% to 111. 9%, and 98. 3% to 115. 1% respectively, while the relative standard deviations ( RSDs) of intra-and inter-assay were below 5%.

doi:10.11895/j.issn.0253-3820.140848

作者:段宏[1]陈雪岚[2]江湖[1]沈骏[1]董胜明[3]熊勇华[1]Andrew Wang[4]

Author:DUAN Hong[1]CHEN Xue-Lan[2]JIANG Hu[1]SHEN Jun[1]DONG Sheng-Ming[3]XIONG Yong-Hua[1]Andrew Wang[4]

作者单位:

 南昌大学食品科学与技术国家重点实验室,南昌,330047

江西师范大学生命科学学院,南昌,330029

北京金源佳和生物科技有限公司,北京,100036

Ocean NanoTech,LLC.,San Diego,CA 92126,USA

刊 名:分析化学 ISTICSCIPKU

Journal:Chinese Journal of Analytical Chemistry年,卷(期):2015, (3)

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量子点荧光微球免疫层析试纸条定量检测恶性疟原虫

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