Yufang Kong1, Huiyu Wang1, Shaoqiang Wu1, Jizhou Lv1, Lin Mei1, Huifang Zhou2, Xiangmei Lin3, Xueqing Han4. 1. Institute of Animal Inspection and Quarantine, Chinese Academy of Inspection and Quarantine, 100176, Beijing, China. 2. People's Hospital of Jiaxiang, Jiaxiang County, Jining City, 272400, Shandong Province, China. 3. Institute of Animal Inspection and Quarantine, Chinese Academy of Inspection and Quarantine, 100176, Beijing, China. linxm@caiq.org.cn. 4. Institute of Animal Inspection and Quarantine, Chinese Academy of Inspection and Quarantine, 100176, Beijing, China. 1422355351@qq.com.
Abstract
BACKGROUND: Brucellosis is a serious zoonosis disease that frequently causes significant economic loss in animal husbandry and threatens human health. Therefore, we established a rapid, accurate, simple and sensitive fluorescent immunochromatographic strip test (ICST) based on quantum dots (QDs) for detection the antibodies of Brucella infection animals serum. RESULTS: The test strips were successfully prepared by quantum dot fluorescent microspheres (QDFM) as tracers, which were covalently coupled to an outer membrane protein of Brucella OMP22. The outer membrane protein OMP28 and monoclonal antibodies of OMP22 were separately dispensed onto a nitrocellulose membrane as test and quality control lines, respectively. The critical threshold for determining negative or positive through the ratio of the fluorescent signal of the test line and the control line (HT / HC) is 0.0492. The repeatability was excellent with an overall average CV of 8.78%. Under optimum conditions, the limit of detection was 1.05 ng/mL (1:512 dilution). With regard to the detection of brucellosis in 150 clinical samples, the total coincidence rate of ICST and Rose Bengal plate test (RBPT) was 97.3%, the coincidence rate of positive samples was 98.8%, the coincidence rate of negative samples was 95.3%, the sensitivity of RBPT is 1:32, and no cross reaction with the sera of other related diseases was observed. CONCLUSION: In our present study, the QDFM has promising application for on-site screening of brucellosis owing to its high detection speed, high sensitivity, high specificity and low cost.
BACKGROUND:Brucellosis is a serious zoonosis disease that frequently causes significant economic loss in animal husbandry and threatens human health. Therefore, we established a rapid, accurate, simple and sensitive fluorescent immunochromatographic strip test (ICST) based on quantum dots (QDs) for detection the antibodies of Brucella infection animals serum. RESULTS: The test strips were successfully prepared by quantum dot fluorescent microspheres (QDFM) as tracers, which were covalently coupled to an outer membrane protein of Brucella OMP22. The outer membrane protein OMP28 and monoclonal antibodies of OMP22 were separately dispensed onto a nitrocellulose membrane as test and quality control lines, respectively. The critical threshold for determining negative or positive through the ratio of the fluorescent signal of the test line and the control line (HT / HC) is 0.0492. The repeatability was excellent with an overall average CV of 8.78%. Under optimum conditions, the limit of detection was 1.05 ng/mL (1:512 dilution). With regard to the detection of brucellosis in 150 clinical samples, the total coincidence rate of ICST and Rose Bengal plate test (RBPT) was 97.3%, the coincidence rate of positive samples was 98.8%, the coincidence rate of negative samples was 95.3%, the sensitivity of RBPT is 1:32, and no cross reaction with the sera of other related diseases was observed. CONCLUSION: In our present study, the QDFM has promising application for on-site screening of brucellosis owing to its high detection speed, high sensitivity, high specificity and low cost.
Authors: Seung Bin Cha; Nabin Rayamajhi; Won Jung Lee; Min Kyung Shin; Myung Hwan Jung; Seung Won Shin; Jong Wan Kim; Han Sang Yoo Journal: FEMS Immunol Med Microbiol Date: 2012-03
Authors: Willem J M Mulder; Karolien Castermans; Judy R van Beijnum; Mirjam G A Oude Egbrink; Patrick T K Chin; Zahi A Fayad; Clemens W G M Löwik; Eric L Kaijzel; Ivo Que; Gert Storm; Gustav J Strijkers; Arjan W Griffioen; Klaas Nicolay Journal: Angiogenesis Date: 2008-12-09 Impact factor: 9.596
Authors: Jeong Ju Lim; Dong Hyeok Kim; Jin Ju Lee; Dae Geun Kim; Wongi Min; Hu Jang Lee; Man Hee Rhee; Hong Hee Chang; Suk Kim Journal: J Vet Med Sci Date: 2011-12-28 Impact factor: 1.267
Authors: Jacques Godfroid; Axel Cloeckaert; Jean-Pierre Liautard; Stephan Kohler; David Fretin; Karl Walravens; Bruno Garin-Bastuji; Jean-Jacques Letesson Journal: Vet Res Date: 2005 May-Jun Impact factor: 3.683
Authors: Paola Caro-Hernández; Luis Fernández-Lago; María-Jesús de Miguel; Ana I Martín-Martín; Axel Cloeckaert; María-Jesús Grilló; Nieves Vizcaíno Journal: Infect Immun Date: 2007-06-11 Impact factor: 3.441