Development of an up-converting phosphor technology-based 10-channel lateral flow assay for profiling antibodies against Yersinia pestis.
In this study, a 10-channel up-converting phosphor technology-based lateral flow (TC-UPT-LF) assay was developed to profile antibodies against Yersinia pestis. Ten expressed Y. pestis proteins were covalently conjugated with an up-converting phosphor particle to develop double-antigen sandwich immunochromatographic strips to detect corresponding antibodies. After optimization one by one, each strip was integrated into a TC-UPT-LF disc for simultaneously detection of different antibodies. A scanning biosensor was also developed to acquire the results. The performance of the TC-UPT-LF assay was evaluated by using standard samples and plague monkey serum samples. Fifty-one patient serum samples were detected by the TC-UPT-LF assay. The TC-UPT-LF disc could be stable for 10 days at 37°C with an average CV of 10.3%. Its sensitivity and qualitative results are comparable to those of ELISA. Its linearity fitting coefficient of determination (R2) for different antibody detection is between 0.93 and 0.99. Besides F1 antibody, the LcrV and YopD antibodies also showed higher positive ratio than the other seven antibodies, as 100% (13/13) and 92% (12/13) in monkey sera and 86.3% (44/51) and 66.7% (34/51) in patient sera, respectively. It is suggested that the TC-UPT-LF assay has been successfully developed for multi-detection and LcrV and YopD can be the potential diagnostic markers of the plague.
Hong
Wenyan
W
Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20, Dongdajie, Fengtai District, Beijing 100071, People's Republic of China.
Huang
Lihua
L
Wang
Haoran
H
Qu
Jianfeng
J
Guo
Zhaobiao
Z
Xie
Chengke
C
Zhu
Ziwen
Z
Zhang
Youbao
Y
Du
Zongmin
Z
Yan
Yanfeng
Y
Zheng
Yan
Y
Huang
Huijie
H
Yang
Ruifu
R
Zhou
Lei
L
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis* / Q000097:blood*; D000942:Antigens, Bacterial; D015374:Biosensing Techniques / Q000379:methods*; D000882:Haplorhini; D006801:Humans; D007118:Immunoassay / Q000379:methods; D010930:Plague / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D015010:Yersinia pestis / Q000276:immunology*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial