摘要
目的对大容量样本中丙型肝炎病毒(HCV)进行浓缩提取,采用自行设计引物进行实时HCV荧光定量检测,验证病毒浓缩的效率以及病毒浓缩对检测的影响。方法采用PEG法高速离心进行病毒浓缩,一步法逆转录荧光定量PCR验证病毒浓缩效果。结果该方案用于HCV RNA检测,能得到良好的扩增曲线,不同病毒载量样本病毒浓缩的效率均在70%以上。结论大容量样本HCV病毒浓缩可提高病毒荧光PCR检测的灵敏度。
Objective The objectives of the present study are to first concentrate and extract Hepatitis C Virus（ HCV） from large samples,and then to identify the efficiency of the virus concentration and its effects on detection by reverse transcription qRT- PCR with custom designed primer. Methods HCV virus concentrated by high speed centrifugation with PEG,and quantitatively determined by one- step RT- PCR.Results This method can be used for HCV RNA test. The results indicated more than 70% concentrating efficiency in samples with different viral load. Conclusion The concentration of HCV virus in large samples can improve the sensitivity of the virus detection by RT- PCR.
出处
《临床和实验医学杂志》
2014年第13期1070-1073,共4页
Journal of Clinical and Experimental Medicine
基金
南京市医学科技发展项目(编号:YKK10131)
