摘要
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 investigate the feasibility of applying nucleic acid extraction-free real-time polymerase chainreaction（PCR） in the cell culture fluid to determine whether the virus is successfully isolated. Methods Dengue and H3 subtype influenza A viruses（IAVs） were isolated from C6/36 and MDCK cells, respectively. Real-time PCR was employed toobtain the Ct values of the extracted nucleic acid and cell culture fluid of the isolated virus, to determine whether the targetpathogen was successfully isolated. Results The Ct values of the nucleic acid extracted from the culture medium of dengueand H3 subtype IAVs ranged from 12.0-14.6 and 8.5-9.8, respectively, while the values of virus culture supernatant were18.5-21.2 and 10.1-12.8, respectively. The qualitative results were perfectly coincident. Conclusion The findings showedthat nucleic acid extraction-free real-time PCR can be directly used in the detection of nucleic acid in the cell culture fluid,with the aim to determine whether the isolation of the target virus is successful or not.
作者
吴衍恒
师舞阳
谢颖
林金思
WU Yanheng SHI Wuyang XIE Ying LIN Jinsi(Zhongshan Center for Disease Control and Prevention, Zhongshan, Guangdong 528400, Chin)
出处
《中国热带医学》
CAS
2017年第1期98-100,共3页
China Tropical Medicine
