Antigen detection based on background fluorescence quenching immunochromatographic assay

Xiangjun Chen, Yangyang Xu, Jinsheng Yu, Jiutong Li, Xuelei Zhou, Chuanyong Wu, Qiuliang Ji, Yuan Ren, Liqun Wang, Zhengyi Huang, Hanling Zhuang, Long Piao, Richard Head, Yajie Wang, Jiatao Lou

Research output: Contribution to journalArticlepeer-review

26 Scopus citations


Gold immunochromatographic assay (GICA) has been around for quite a while, but it is qualitative in the vast majority of applications. A fast, simple and quantitative GICA is in call for better medicine. In the current study, we have established a novel, quantitative GICA based on fluorescence quenching and nitrocellulose membrane background signals, called background fluorescence quenching immunochromatographic assay (bFQICA). Using model analyte alpha-fetoprotein (AFP), the present study assessed the performance of bFQICA in numerous assay aspects. With serial dilutions of the international AFP standard, standard curves for the calculation of AFP concentration were successfully established. At 10 and 100ngmL-1 of the international AFP standard, the assay variability was defined with a coefficient of variance at 10.4% and 15.2%, respectively. For samples with extended range of AFP levels, bFQICA was able to detect AFP at as low as 1ngmL-1. Fluorescence in bFQICA strips stayed constant over months. A good correlation between the results from bFQICA and from a well-established Roche electrochemiluminescence immunoassay was observed in 27 serum samples (r=0.98, p<0.001). In conclusion, our study has demonstrated distinctive features of bFQICA over conventional GICA, including utilization of a unique fluorescence ratio between nitrocellulose membrane background and specific signals (F1/F2) to ensure accurate measurements, combined qualitative and quantitative capabilities, and exceptionally high sensitivity for detection of very low levels of antigens. All of these features could make bFQICA attractive as a model for antigen-antibody complex based GICA, and could promote bFQICA to a broad range of applications for investigation of a variety of diseases.

Original languageEnglish
Pages (from-to)44-50
Number of pages7
JournalAnalytica Chimica Acta
StatePublished - Sep 2 2014


  • Background fluorescence quenching
  • Immunochromatography
  • Nitrocellulose membrane
  • Point of care test
  • Quantitative detection


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