超声波均质化对仙台病毒ELISA测试中抗原稳定性的影响

目的研究超声波均质化(homogenization)处理对于仙台病毒在ELISA测试中抗原稳定性的影响。方法对BHK-21细胞内扩增培养的仙台病毒通过差速离心,富集后使用超声波做均质化处理,和未处理的病毒分别包被酶标板,使用标准免疫小鼠血清和SPF小鼠血清对包被平板进行测试,比较样品测试孔间测量数据的变异率。结果对免疫血清做梯度稀释,各梯度样品在未经超声处理仙台病毒抗原包被ELISA检测中测量值的变异率在1.97%～6.02%之间;在经超声处理仙台病毒抗原包被ELISA检测中测量值的变异率在0.53%～2.26%之间;SPF血清测量值的变异率均高于免疫血清;所有样品在经超声处理的病毒抗原包被ELISA检测中测量值的变异率均较小。结论超声波处理有效的提高了仙台病毒抗原的均质性,在ELISA测试中提高了抗原的稳定性。

Ultrasonic Homogenization Improves the Sendai Virus Antigen Stability in ELISA

Objective To analyze the effects of ultrasonic homogenization on the stability of Sendai virus(SeV) in ELISA.Methods The SeV amplified in BHK-21 cells was purified by differential centrifugation,and then homogenized by ultrasonication.The ultrasonic-treated SeV antigen and the untreated antigen were coated in the wells of ELISA plates.The coefficients of variation(CV) between the two kinds of plates were compared by testing SPF mouse serum and SeV-immunized serum.Results the CV of gradient diluted Sendai virus-immunized serum was between 1.97% to 6.02% in untreated antigen coated plates and 0.53% to 2.26% in ultrasonic-treated antigen coated plates.The CV of SPF sera was higher than that of SeV-immunized serum.The CV of all the tested samples in ultrasonic-treated antigen coated plates was lower than that in the untreated antigen coated plates.Conclusion Ultrasonic treatment can homogenize the SeV antigen and improve the SeV antigen stability in ELISA.