Abstract:The aim of this study was to compare three kinds of commercial enzyme-linked immunosorbent assay (ELISA) kits for detection of antibodies against Japanese encephalitis virus (JEV) in swine sera. A total of 90 swine serum samples were tested simultaneously by three kinds of commercial JEV IgG ELISA kits (Keqian, Lvshiyuan and Tianchen), which were used with the most frequently recorded in domestic literatures. Diagnostic test methods were used for evaluatlon. Neutralization test was used as the gold standard’ test. Results showed that all the three kinds of commercial ELISA kits had higher positive detection rate than that of neutralization test (34.4%, 31/90), the rates of Keqian, Lvshiyuan and Tianchen were 50.0% (45/90), 47.8% (43/90) and 38.9% (35/90), respectively. Both of Lvshiyuan and Tianchen had a result of suspicious positive, the rates were 32.2% (29/90) and 4.4% (4/90) respectively. Sensitivity and specificity of Tianchen was 83.87% and 79.66%, respectively, with the highest positive likelihood ratio (4.12); Keqian had the highest sensitivity (90.32%) and a lower specificity (71.19%), with the highest negative likelihood ratio (0.14); while Lvshiyuan had the lowest sensitivity and specificity, which was 41.94% and 16.95%, respectively. There was no significant difference of detected rates between Tianchen and the gold standard’ serum neutralization test with the highest goodness of fit (κ = 0.63). Comparison results of Keqian and Lvshiyuan with neutralization test were significantly different: the κ coefficient was 0.57 and 0.04, respectively. Additionally, internal consistency analysis suggested that all the ELISA kits had good stability. In conclusion, currently commercially available ELISA kits for detection of JEV IgG antibody in swine sera are quite various. A high false negative was still found in these kits compared with neutralization test. Our results suggest the need to improve the insure of authenticity and reliability in use of commercial JEV IgG ELISA kits.
[1] Ze WY.Plan immunology[M]. 2nd ed. Shanghai: Shanghai Science and Technology Literature Publishing House, 2002,28-53. (in Chinese) 迮文远.计划免疫学.2版[M]. 上海:上海科技文献出版社,2002,28-53. [2] Song DW, He SH. A review of immunoassays in the serodiagnosis of Japanese encephalitis virus[J]. J Agr Sci, 2006, 27(1): 72-74. (in Chinese) 宋大伟,何生虎. 乙型脑炎血清学诊断方法研究进展[J]. 农业科学研究,2006,27(1):72-74. [3] Ting SH, See E, Tan HC, et al. Development of a simplified assay for the detection of neutralizing antibodies to Japanese encephalitis virus[J]. J Virol Methods, 2001, 93(1/2): 43-47. doi:10.1016/S0166-0934(00)00293-7 [4] Hamano M, Lim CK, Takagi H, et al. Detection of antibodies to Japanese encephalitis virus in the wild boars in Hiroshima prefecture, Japan[J]. Epidemiol Infect, 2007, 135(6): 974-977. doi:10.1017/S0950268806007710 [5] Chen XM, Qi WB, Zhang GH, et al. Monitoring and analysis of antibodies against Japanese encephalitis virus in part of large-scale farms in Guangdong Province[J]. Guangdong Agr Sci, 2010, 37(9): 154-155. (in Chinese) 陈孝明,亓文宝,张桂红,等. 广东省部分规模化猪场乙脑抗体的监测与分析[J]. 广东农业科学,2010,37(9):154-155. [6] Ma HB, Tan H, Sun H, et al. Sero-epidemiological survey on Japanese encephalitis at pig farms registered in Zhuhai[J]. Chin J Frontier Health Quarantine, 2007, 30(3): 142-144. (in Chinese) 马洪波,谭华,孙虹,等. 珠海市部分注册猪场日本脑炎调查研究[J]. 中国国境卫生检疫杂志,2007,30(3):142-144. [7] Yang RF, Bi ZX. Epidemiological survey on Japanese encephalitis in pigs in Shandong Province[J]. Anim Husb Vet Med, 2008, 40(10): 61-62. (in Chinese) 杨瑞锋,毕志香. 山东省猪日本脑炎流行病学调查[J]. 畜牧与兽医,2008,40(10):61-62. [8] Cheng ZT, Chen JY, Yue J, et al. The sero-epidemiological survey of porcine epidemic encephalitis B in Guizhou Province[J]. Chin J Zoonoses, 2011, 27(12): 1156-1158. (in Chinese) 程振涛,陈军义,岳筠,等. 贵州省猪乙型脑炎血清流行病学调查[J]. 中国人兽共患病学报,2011,27(12):1156-1158. [9] Qiao JP, Zhao MQ, Zhang XT, et al. Preparation and comparison of three dfferent adjuvant inactivated vaccines of Japanese encephalitis virus from swine[J]. J South China Agr Univ, 2011, 32(2): 85-88. (in Chinese) 乔进平,赵明秋,张学涛,等. 猪乙型脑炎病毒3种灭活疫苗的制备及免疫效果比较[J]. 华南农业大学学报,2011,32(2):85-88. [10] Shen T, Zhou B, Chen PY. Development and application of an indirect ELISA method for detection of antibody to porcine Japanese encephalitis virus[J]. Anim Husb Vet Med, 2010, 42(4): 1-7. (in Chinese) 沈婷,周斌,陈溥言. 检测猪乙型脑炎病毒抗体间接ELISA方法的建立与应用[J]. 畜牧与兽医,2010,42(4):1-7. [11] Chen ZS, Wang EQ, Zhang H, et al. Evaluation of pig live attenuated Japanese encephalitis vaccines in a farm of Baoding[J]. Heilongjiang Anim Sci Vet Med, 2011(07): 121-122. (in Chinese) 陈振师,汪恩强,张恒,等. 保定市某猪场乙型脑炎减毒活疫苗免疫效果评价[J]. 黑龙江畜牧兽医,2011(07):121-122. [12] Lindahl J, Boqvist S, Stahl K, et al. Reproductive performance in sows in relation to Japanese encephalitis virus seropositivity in an endemic area[J]. Trop Anim Health Prod, 2012, 44(2): 239-245. doi:10.1007/s11250-011-0005-0 [13] Huang YH, Yin ZX, Xie GH, et al. Porcine epidemiological survey on Japanese encephalitis in Dongguan in Guangdong Province[J]. Progr Vet Med, 2012, 33(7): 126-128. (in Chinese) 黄育浩,殷自兴,谢国怀,等. 广东省东莞市猪乙型脑炎流行病学调查[J]. 动物医学进展,2012,33(7):126-128. [14] Hombach J, Solomon T, Kurane I, et al. Report on a WHO consultation on immunological endpoints for evaluation of new Japanese encephalitis vaccines, WHO, Geneva, 2-3 September, 2004[J]. Vaccine, 2005, 23(45): 5205-5211. doi:10.1016/j.vaccine.2005.07.002 [15] Gao XY, Fu SH, Wang HY, et al. Establishment of IgG capturing E-linked immunosorbent assay to detect Japanese encephalitis virus antibody[J]. Dis Surveill, 2010, 25(2): 107-110, 119. (in Chinese) 高晓艳,付士红,王环宇,等. 流行性乙型脑炎病毒IgG抗体捕获酶联免疫吸附试验检测方法的建立[J]. 疾病监测,2010,25(2):107-110, 119.