To evaluate Mycoplasma detecting methods of lyophilized porcine reproductive and respiratory syndrome virus (PRRSV) vaccine productions, three methods which included DNA fluorescence staining, PCR and virus isolation for the Mycoplasma detections of the lyophilized PRRSV semi- and finished products. DNA fluorescence staining was suitable method in detecting Mycoplasma contamination for batches of the vaccine semi-products in 6 days. Ten batches of products were detected and the results showed that DNA fluorescence staining and PCR were reliable methods in detecting Mycoplasma, and PCR was more sensitive than the other 2 methods. DNA fluorescence staining and PCR were easy to use and less cost. Therefore, both methods could be used for the quality control of the vaccine production procedure.%为完善猪繁殖与呼吸综合征病毒(PRRSV)冻干疫苗生产各阶段的支原体检测体系,本研究分别应用DNA荧光染色法、PCR法和病毒分离培养法检测疫苗生产中的细胞、种毒、半成品、成品,分析方法的可行性和优势.结果显示,DNA荧光染色法检测PRRSV半成品抗原液的支原体污染能够于6d内得到结果,与其余半成品检验项目用时相近,适宜在生产中应用.而采用DNA荧光染色、PCR和病毒分离培养法对我们生产的10批PRRSV冻干疫苗成品的支原体检测结果显示,DNA荧光染色法与病毒分离培养法检测的结果一致,1批疫苗的PCR检测结果与培养法检测结果不符,即PCR法检测阳性,但DNA荧光染色和病毒分离培养法检测为阴性.由此证明,DNA荧光染色法和PCR法检测疫苗中支原体的结果可靠,而PCR法检出率更高.DNA荧光染色法和PCR法操作简便、快速、经济,可以作为疫苗生产质量的内控标准,提高支原体的检出率,保证疫苗质量.
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