.2006 Dec 7;119(2):656-663.

doi: 10.1016/j.snb.2006.01.029. Epub 2006 Feb 17.

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

Zhongqiang Yan¹, Lei Zhou¹, Yongkai Zhao², Jing Wang³, Lihua Huang², Kongxin Hu³, Haihong Liu¹, Hong Wang¹, Zhaobiao Guo¹, Yajun Song¹, Huijie Huang², Ruifu Yang¹

Affiliations

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.

PMID:32288237
PMCID: PMC7125792
DOI: 10.1016/j.snb.2006.01.029

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

Zhongqiang Yan et al. Sens Actuators B Chem.2006.

.2006 Dec 7;119(2):656-663.

doi: 10.1016/j.snb.2006.01.029. Epub 2006 Feb 17.

Authors

Zhongqiang Yan¹, Lei Zhou¹, Yongkai Zhao², Jing Wang³, Lihua Huang², Kongxin Hu³, Haihong Liu¹, Hong Wang¹, Zhaobiao Guo¹, Yajun Song¹, Huijie Huang², Ruifu Yang¹

Affiliations

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.

PMID:32288237
PMCID: PMC7125792
DOI: 10.1016/j.snb.2006.01.029

Abstract

Up-converting phosphor technology (UPT)-based lateral-flow immunoassay has been developed for quantitative detection ofYersinia 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 ofY. 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 andV_C stand for the multiplied voltage units for the test and the control line, respectively, and the ratioV_T/V_C is directly proportional to the number ofY. pestis in a sample. We observed a good linearity between the ratio and log CFU/ml ofY. 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.

Keywords: Lateral-flow immunoassay; Sandwich immunoassay; Up-converting phosphor technology; Yersinia pestis.

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Figures

**Fig. 1**
Schematic description of the UPT-based lateral-flow test strip. The components of the strip include a sample pad (1), a conjugate pad (2), a nitrocellulose membrane (3), an absorbent pad (4), and a laminating card (5). During the manufacture, a test line (6), a control line (7), and an ending index line (8) are disposed on the strip and other components are pasted on the laminating card with proper overlapping.

**Fig. 2**
Photograph of the portable prototype UPT-based biosensor.

**Fig. 3**
Schematic representation of the design of UPT-based biosensor.

**Fig. 4**
The illustrations of UPT-based lateral-flow immunoassay test results.

**Fig. 5**
The dose-response curve obtained from the mean ratioV_T/V_C against log (*Y. pestis* EV76 CFU/ml) in the dilution buffer. Six repeated tests of each concentration were performed.

**Fig. 6**
Correlation between the ratioV_T/V_C and the log (*Y. pestis* EV76 CFU/ml) in the dilution buffer. Six repeated tests of each concentration were performed.

**Fig. 7**
Correlation between the UPT-LF and the plate counting forY. pestis strain 201 in lung samples of the experimentally infected Balb/c mice.

See this image and copyright information in PMC

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

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

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