Lateral Flow Immunoassay Using Europium Chelate-Loaded Silica Nanoparticles as Labels

被引:106
作者
Xia, Xiaohu [2 ]
Xu, Ye [2 ]
Zhao, Xilin [2 ,3 ]
Li, Qingge [1 ,2 ,4 ]
机构
[1] Xiamen Univ, Sch Life Sci, Dept Biomed Sci, Mol Diagnost Lab, Xiamen 361005, Fujian, Peoples R China
[2] Xiamen Univ, Sch Life Sci, Key Lab Minist Educ Cell Biol & Tumor Cell Engn, Xiamen 361005, Fujian, Peoples R China
[3] Univ Med & Dent New Jersey, Publ Hlth Res Inst, Newark, NJ USA
[4] Xiamen Univ, Key Lab Chem Biol Fujian, Xiamen 361005, Fujian, Peoples R China
关键词
RESOLVED IMMUNOFLUOROMETRIC ASSAYS; PROSTATE-SPECIFIC ANTIGEN; ULTRASENSITIVE DETECTION; LIPOSOME IMMUNOASSAY; AFLATOXIN B-1; CHROMATOGRAPHY; FLUORESCENCE; SYSTEM;
D O I
10.1373/clinchem.2008.114561
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
BACKGROUND: Despite their ease of use, lateral flow immunoassays (LFIAs) often suffer from poor quantitative discrimination and low analytical sensitivity. We explored the use of a novel class of europium chelate-loaded silica nanoparticles as labels to overcome these limitations. METHODS: Antibodies were covalently conjugated onto europium chelate-loaded silica nanoparticles with dextran as a linker. The resulting conjugates were used as labels in LFIA for detection of hepatitis B surface antigen (HBsAg). We performed quantification with a digital camera and Adobe Photoshop software. We also used 286 clinical samples to compare the proposed method with a quantitative ELISA. RESULTS: A detection limit of 0.03 mu g/L was achieved, which was 100 times lower than the colloidal gold-based LFIAs and lower than ELISA. A precise quantitative dose-response curve was obtained, and the linear measurement range was 0.05-3.13 mu g/L, within which the CVs were 2.3%-10.4%. Regression analysis of LFIA on ELISA results gave: log (LFIA) = -0.14 log (ELISA) + 1.03 mu g/L with r = 0.99 for the quantification of HBsAg in 35 positive serum samples. Complete agreement was observed for the qualitative comparison of 286 clinical samples assayed with LFIA and ELISA. CONCLUSIONS: Europium chelate-loaded silica nano-particle labels have great potential to improve LFIAs, making them useful not only for simple screening applications but also for more sensitive and quantitative immunoassays.
引用
收藏
页码:179 / 182
页数:4
相关论文
共 17 条
[1]   Ganglioside-liposome immunoassay for the ultrasensitive detection of cholera toxin [J].
Ahn-Yoon, S ;
DeCory, TR ;
Baeumner, AJ ;
Durst, RA .
ANALYTICAL CHEMISTRY, 2003, 75 (10) :2256-2261
[2]   LATEX-BASED THIN-LAYER IMMUNOAFFINITY CHROMATOGRAPHY FOR QUANTITATION OF PROTEIN ANALYTES [J].
BIRNBAUM, S ;
UDEN, C ;
MAGNUSSON, CGM ;
NILSSON, S .
ANALYTICAL BIOCHEMISTRY, 1992, 206 (01) :168-171
[3]   An enzyme immunoanalytical system based on sequential cross-flow chromatography [J].
Cho, JH ;
Paek, EH ;
Cho, IH ;
Paek, SH .
ANALYTICAL CHEMISTRY, 2005, 77 (13) :4091-4097
[4]   Development of an immunoassay-based lateral flow dipstick for the rapid detection of aflatoxin B1 in pig feed [J].
Delmulle, BS ;
De Saeger, SMDG ;
Sibanda, L ;
Barna-Vetro, I ;
Van Peteghem, CH .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2005, 53 (09) :3364-3368
[5]   One-step immunostrip test for the simultaneous detection of free and total prostate specific antigen in serum [J].
Fernández-Sánchez, C ;
McNeil, CJ ;
Rawson, K ;
Nilsson, O ;
Leung, HY ;
Gnanapragasam, V .
JOURNAL OF IMMUNOLOGICAL METHODS, 2005, 307 (1-2) :1-12
[6]   Upconverting phosphor reporters in immunochromatographic assays [J].
Hampl, J ;
Hall, M ;
Mufti, NA ;
Yao, YMM ;
MacQueen, DB ;
Wright, WH ;
Cooper, DE .
ANALYTICAL BIOCHEMISTRY, 2001, 288 (02) :176-187
[7]  
Härmä H, 2001, CLIN CHEM, V47, P561
[8]   EUROPIUM AS A LABEL IN TIME-RESOLVED IMMUNOFLUOROMETRIC ASSAYS [J].
HEMMILA, I ;
DAKUBU, S ;
MUKKALA, VM ;
SIITARI, H ;
LOVGREN, T .
ANALYTICAL BIOCHEMISTRY, 1984, 137 (02) :335-343
[9]   A strip liposome immunoassay for aflatoxin B1 [J].
Ho, JAA ;
Wauchope, RD .
ANALYTICAL CHEMISTRY, 2002, 74 (07) :1493-1496
[10]   Development of an ultrarapid one-step fluorescence immunochromatographic assay system for the quantification of microcystins [J].
Kim, YM ;
Oh, SW ;
Jeong, SY ;
Pyo, DJ ;
Choi, EY .
ENVIRONMENTAL SCIENCE & TECHNOLOGY, 2003, 37 (09) :1899-1904