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Rapid quantitative detection of Yersinia pestis by lateral-flow immunoassay and up-converting phosphor technology-based biosensor

Authors
  • Yan, Zhongqiang1
  • Zhou, Lei1
  • Zhao, Yongkai2
  • Wang, Jing3
  • Huang, Lihua2
  • Hu, Kongxin3
  • Liu, Haihong1
  • Wang, Hong1
  • Guo, Zhaobiao1
  • Song, Yajun1
  • Huang, Huijie2
  • Yang, Ruifu1
  • 1 Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, National Center for Biomedical Analysis, Army Center for Microbial Detection and Research, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences (AMMS), Beijing 100071, PR China
  • 2 Shanghai Institute of Optics and Fine Mechanics (SIOM), Chinese Academy of Sciences, Shanghai 201800, PR China
  • 3 National Academy of Inspection and Quarantine, Beijing 100025, PR China
Type
Published Article
Journal
Sensors and Actuators B Chemical
Publisher
Elsevier
Publication Date
Feb 17, 2006
Volume
119
Issue
2
Pages
656–663
Identifiers
DOI: 10.1016/j.snb.2006.01.029
PMID: 32288237
PMCID: PMC7125792
Source
PubMed Central
Keywords
License
Unknown

Abstract

Up-converting phosphor technology (UPT)-based lateral-flow immunoassay has been developed for quantitative detection of Yersinia pestis rapidly and specifically. In this assay, 400 nm up-converting phosphor particles were used as the reporter. A sandwich immumoassay was employed by using a polyclonal antibody against F1 antigen of Y. pestis immobilized on the nitrocellulose membrane and the same antibody conjugated to the UPT particles. The signal detection of the strips was performed by the UPT-based biosensor that could provide a 980 nm IR laser to excite the phosphor particles, then collect the visible luminescence emitted by the UPT particles and finally convert it to the voltage as a signal. V T and V C stand for the multiplied voltage units for the test and the control line, respectively, and the ratio V T/ V C is directly proportional to the number of Y. pestis in a sample. We observed a good linearity between the ratio and log CFU/ml of Y. pestis above the detection limit, which was approximately 104 CFU/ml. The precision of the intra- and inter-assay was below 15% (coefficient of variation, CV). Cross-reactivity with related Gram-negative enteric bacteria was not found. The UPT-LF immunoassay system presented here takes less than 30 min to perform from the sample treatment to the data analysis. The current paper includes only preliminary data concerning the biomedical aspects of the assay, but is more concentrated on the technical details of establishing a rapid manual assay using a state-of-the-art label chemistry.

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