大鼠仙台病毒ELISA、间接免疫荧光和免疫印迹三种检测方法比较

A comparison of rat sendai virus detection methods of ELISA,IFA and WB

目的比较ELISA(enzyme-linked immunosorbent assay)、IFA(immuno-fluorescence assay)和WB(Western blot)三种方法在大鼠仙台病毒血清学检测中的差异。方法仙台病毒蛋白抗原经凝胶电泳分离转移后用于血清学检测的WB方法;使用IFA、ELISA方法对20份无菌大鼠、227份SPF大鼠以及63份清洁级大鼠送检血清样品进行检测,阳性及可疑样品用WB方法进行了验证。结果 20份无菌大鼠血清样品被3种方法检测为仙台病毒抗体阴性;SPF级大鼠样品被IFA方法判定为阴性,1.32%(3/227)被ELISA方法判定为阳性,其中有2/3被WB确认为阳性;ELISA、IFA和WB在清洁级大鼠样品中检出仙台病毒的阳性率分别为为18.12%、11.34%和15.87%。结论三种检测方法灵敏度从高到低依次为ELISA、WB和IFA。WB方法可作为IFA和ELISA难以确定结果的替代方法。

Objective to compare the three methods of ELISA, IFA and WB in the detection of Sendai virus in rat sera. Methods the SeV antigen proteins separated by SDS-PAGE were used in the WB method; 20 sera from germ free rats, 227 sera from SPF rats and 63 sera from clean rats were analyzed by ELISA, IFA,those candidate positive samples were tested by WB. Results The 20 germ free rats were determined SeV negative by all of the 3 methods; all of the SPF rats were determined SeV negative by IFA, but 1.32% of these SPF rats were determined Se~ positive by ELISA, and 2/ 3 of these ELISA determined positive sera were confrrmed by WB; The SeV positive rate in clean rats were 18.12% by ELISA, 11.34% by IFA, and 15.87% by WB. Conclusion thedetection sensitivity gradient of the 3 methods from high to low is ELISA, WB, and IFA. And WB should be an alternative SeV detection method for those indeterminate samplvs by IFA and ELISA.