Comparative study of FPA and ELISA method in the diagnosis of animal brucellosis
REN Yan-bin1, 2, ZHANG Hui2, SUN Xiang-xiang2, LI Ze-qin3, CAI Jia-li1, FAN Wei-xing3
1. Chongqing University of Technology, Chongqing 404100, China; 2. China Animal Health and Epidemiology Center, Qingdao 266000, China; 3. Pinghe Biotechnology Institute, Heilongjiang 150000, China
Abstract:In order to evaluate the application of fluorescence polarization test (FPA) for clinical detection of serum sample, 1 286 samples of cattle serum which included 309 positive sera and 977 negative serum samples were tested, and then we used the ROC curve to analyse the FPA critical value. The sensitivity and specificity of FPA, IELISA, and CELISA were compared through testing 1 160 samples of cattle and analyzing, and the consistency of these three detection methods were applied by using Kappa analysis. The result of the analysis of ROC curve showed that 95 mP was the optimal threshold for the test of FPA; the diagnostic sensitivity and specificity were 100% and 99.6% respectively; the sensitivity were 97%, 99.56% and 98.27% respectively; the specificity were 100%, 98.06%, and 99.56% for the diagnostic of FPA, IELISA and CELISA respectively. At the same time, consistency of the three methods was all≥0.75. FPA could be used in animal brucellosis diagnosis under field conditions. The rapid and accurate diagnosis of brucellosis could avoid animal twice arrested and suitable for application.
任燕斌, 张慧, 孙翔翔, 李泽钦, 蔡家利, 范伟兴. FPA与ELISA方法在我国部分地区牛布鲁菌病诊断应用中的比较研究[J]. 中国人兽共患病学报, 2016, 32(9): 806-809.
REN Yan-bin, ZHANG Hui, SUN Xiang-xiang, LI Ze-qin, CAI Jia-li, FAN Wei-xing. Comparative study of FPA and ELISA method in the diagnosis of animal brucellosis. Chinese Journal of Zoonoses, 2016, 32(9): 806-809.
[1] Godfroid J, Nielsen K, Saegerman C. Diagnosis of brucellosis in livestock and wildlife[J]. Croat Med J, 2010, 51: 296-305. [2] Peng AY, Zhang YC, Wang WJ. The comparison of brucellosis detection method for livestock[J]. China Anim Husband Vet Abstracts, 2013, 29(3): 48. (in Chinese) 彭安业,张迎春,王文娟. 家畜布氏杆菌检测方法比较[J]. 中国畜牧兽医文摘, 2013,29(3): 48. [3] Zhong YM, Sun G, Zhang JF, et al. The comparison between IELISA and normal method for detection of bovine brucellosis[J]. Chin J Ctrl Endem Dis, 2009, 24(1): 36-37. (in Chinese) 钟映梅,孙刚,张俊峰,等. 应用间接酶联免疫吸附试验方法与常规方法诊断牛、羊布鲁菌病的比较试验[J]. 中国地方病防治杂志,2009,24(1):36-37. [4] Qi JW, Yi YX, Zhou YH, et al. Comparative analysis of four kinds of serological detection methods for bovine brucelosis and belitensis brucellosis[J]. Chin J Anim Quarant, 2015, 32(4): 57-59. (in Chinese) 齐景文, 依颖新, 周艳红, 等. 牛羊布鲁菌病四种血清学检测方法对比分析[J]. 中国动物检疫. 2015,32(4):57-59. [5] OIE. OIE Terrestrial Manual[M]. Beijing:China Agriculture Press, 2012:616-650. 世界动物卫生组织. 陆生动物卫生法典[M]. 北京:中国农业出版社, 2012:616-650. [6] Nielsen K, Gall D, Jolley M, et al. A homogeneous fluorescence polarization assay for detection of antibody to Brucella abortus [J]. J Immunol Methods, 1996, 195: 161-168. doi:10.1016/0022-1759(96)00116-0 [7] Sanchez-Villalobos A, Urdaneta-Fernandez M, Rubio-Fuenmayor E, et al. Development and evaluation of a serological protocol of fluorescence polarization for the preliminary study of Brucella spp. antibodies in humans[J]. Invest Clin, 2011, 52: 48-57. [8] Lynch M, Duignan PJ, Taylor T, et al. Epizootiology of Brucella infection in Australian fur seals[J]. J Wildl Dis, 2011, 47: 352-363. doi:10.7589/0090-3558-47.2.352 [9] Nicola AM, Elena S, Alonso B, et al. Evaluation of the Fluorescence Polarization Assay (FPA) for diagnosis of Brucella melitensis infection of goats in Argentina[J]. Prilozi, 2010, 31: 133-143. [10] Her M, Cho DH, Kang SI, et al. Outbreak of brucellosis in domestic elk in Korea[J]. Zoonoses Public Hlth, 2010, 57: 155-161. doi:10.1111/j.1863-2378.2009.01287.x [11] Nielsen K, Smith P, Yu W, et al. Towards single screening tests for brucellosis[J]. Rev Sci Tech, 2005, 24: 1027-1037. [12] Minas A, Stournara A, Minas M, et al. Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B. melitensis infection in sheep and goats[J]. J Immunol Methods, 2007, 320: 94-103. doi:10.1016/j.jim.2006.12.008 [13] Nielsen K, Smith P, Gall D, et al. Validation of the fluorescence polarization assay for detection of milk antibody to Brucella abortus [J]. J Immunoassay Immunochem, 2001, 22: 203-211. doi:10.1081/IAS-100104706 [14] Nielsen K, Gall D, Smith P, et al. Fluorescence polarization assay for the diagnosis of bovine brucellosis: adaptation to field use[J]. Vet Microbiol, 2001, 80: 163-170. doi:10.1016/S0378-1135(00)00386-2 [15] Wang YL, Wang K, Zhang JZ, et al. Comparison on the clinical application of six serological diagnosis methods for Brucella [J]. Chin J Frontier Hlth Quarant, 2014, 37(5): 318-320. (in Chinese) 王玉玲, 王可, 张俊哲, 等. 布鲁菌6种血清学检测方法的临床应用比较[J]. 中国国境卫生检疫杂志,2014,37(5):318-320. [16] Wang XL, Jiang YW, Mao KR, et al. Research progress of laboratory diagnostic methods for brucellosis[J]. Chin J Vet Drug, 2011, 45(11): 37-42. (in Chinese) 王秀丽,蒋玉文,毛凯荣,等. 布鲁菌病实验室诊断方法的研究进展[J]. 中国兽药杂志, 2011, 45(11):37-42. [17] Gall D, Nielsen K, Forbes L. Validation of the fluorescence polarization assay and comparison to other serological assays for the detection of serum antibodies to Brucella abortus in bison[J]. J Wldlife Dis, 2000, 36(3): 469-476. doi:10.7589/0090-3558-36.3.469 [18] Liu XM, Yang H, Gao MC, et al. The Bayesian assessment of three serological diagnostic methods for brucellosis[J]. Anim Husband Vet Sci, 2010, 41(4): 449-453. (in Chinese) 刘晓民,杨华,高明春,等. 3种奶牛布鲁菌病血清学检测方法的贝叶斯评估[J]. 畜牧兽医学报, 2010,41(4):449-453.