Quantitative detection in the attomole range for immunochromatographic tests by means of a flatbed scanner

被引:69
作者
Lönnberg, M
Carlsson, J
机构
[1] Pharmacia Diagnost AB, S-75182 Uppsala, Sweden
[2] Univ Uppsala, Ctr Surface Biotechnol, S-75123 Uppsala, Sweden
关键词
carbon black; desktop scanner; flatbed scanner; IgE; immunoassay; immunochromatography; lateral flow test; point-of-care testing;
D O I
10.1006/abio.2001.5130
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This work describes the use of the combination of carbon black as an antibody label, a membrane-based immunochromatographic device, and a flatbed scanner as a quantitative test system. The scanner detected 0.4-345 ng carbon black/mm(2) on a nitrocellulose membrane (0.2-170 amol carbon black/mm2) with an imprecision (coefficient of variation, CV) lower than 2% for the carbon black determination and a detection limit of 0.04 ng carbon black/mm(2) (0.02 amol/mm(2)), The detection ability was compared to that obtained with alkaline phosphatase (ALP) using a substrate yielding a chemiluminescent signal (0.02 amol ALP/well), beta -galactosidase using a substrate yielding a fluorescent signal (0.3 amol beta -galactosidase/well), and horseradish peroxidase (HRP) using a substrate yielding a colored signal (5 amol HRP/microtiter well). The carbon black immunochromatographic test for immunoglobulin E (IgE) showed a detection limit of 0.13 pM IgE (0.01 kU/L) after a testing time of 10 min. The scanner detection imprecision for the IgE determination was 0.6% CV in the range 1-10 kU IgE/L when 2.3 mm(2) was used for detection and 1% CV when 0.19 mm2 was used. A flatbed scanner is an inexpensive instrument with multiple uses, which now also includes the sensitive evaluation of immunoassays, (C) 2001 Academic Press.
引用
收藏
页码:224 / 231
页数:8
相关论文
共 9 条
  • [1] Corrie, 1983, IMMUNOASSAYS CLIN CH, P76
  • [2] DONOVAN RS, 1994, BIOTECHNIQUES, V17, P660
  • [3] KUHNER G, 1992, PT171530992
  • [4] Membrane assisted isoform immunoassay -: A rapid method for the separation and determination of protein isoforms in an integrated immunoassay
    Lönnberg, M
    Carlsson, J
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 2000, 246 (1-2) : 25 - 36
  • [5] Müller-Bardorff M, 1999, CLIN CHEM, V45, P1002
  • [6] PRICE CP, 1997, PRINCIPLES PRACTISE, P580
  • [7] Scanometric DNA array detection with nanoparticle probes
    Taton, TA
    Mirkin, CA
    Letsinger, RL
    [J]. SCIENCE, 2000, 289 (5485) : 1757 - 1760
  • [8] QUANTITATIVE COMPUTER IMAGE-ANALYSIS OF A HUMAN CHORIONIC-GONADOTROPIN COLLOIDAL CARBON DIPSTICK ASSAY
    VANAMERONGEN, A
    VANLOON, D
    BERENDSEN, LBJM
    WICHERS, JH
    [J]. CLINICA CHIMICA ACTA, 1994, 229 (1-2) : 67 - 75
  • [9] VELLEMAN SG, 1995, BIOTECHNIQUES, V18, P1056