论文部分内容阅读
目的探讨应用免核酸提取Real-time PCR技术直接对细胞培养液进行检测鉴定,判断成功分离病毒的可行性。方法应用C6/36细胞和MDCK细胞分别对登革病毒和甲型H3亚型流感病毒进行病毒分离培养,应用Real-timePCR技术分别对经提取的核酸溶液和细胞培养液进行目标病原体检测,判断是否成功分离病毒。结果登革病毒和甲型H3亚型流感病毒细胞培养液经核酸提取后进行Real-time PCR检测,检测结果CT值范围分别为12.0~14.6和8.5~9.8,病毒培养液直接检测结果范围分别为18.5~21.2和10.1~12.8,定性结果完全一致。结论免核酸提取Real-timePCR技术可直接对细胞培养液进行病毒核酸检测,判断是否成功分离病毒。
Objective To explore the feasibility of using real-time PCR-free nucleic acid to directly detect and identify cell culture liquid to determine the successful virus isolation. Methods C6 / 36 cells and MDCK cells were respectively used for virus isolation and culture of dengue virus and influenza A (H3) virus. Real-time PCR was used to detect the target pathogens in the extracted nucleic acid solution and cell culture medium, respectively, to determine whether Successfully isolated virus. Results Real-time PCR detection of dengue virus and influenza A virus subtype H3 influenza virus was carried out after nucleic acid extraction. The CT values ranged from 12.0 to 14.6 and 8.5 to 9.8, respectively. The direct detection results of the virus culture solution were 18.5 ~ 21.2 and 10.1 ~ 12.8, the qualitative results are exactly the same. Conclusion Nucleic acid-free extraction Real-timePCR technology can be directly on the cell culture medium for nucleic acid detection to determine whether the successful virus isolation.