The comparison of three different nucleic acid isolation methods in hepatitis B virus DNA detection with real-time quantitative PCR
- VernacularTitle:三种HBV DNA提取方法对荧光定量PCR检测结果影响的比较
- Author:
Jio LIU
;
Jun XU
;
Xuefei WANG
;
Haibin WANG
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus;
DNA,viral;
Polymerase chain reaction
- From:
Chinese Journal of Laboratory Medicine
2008;31(7):780-783
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the influence of different HBV DNA extraction methods with Micro-Nucleic-Releaser (MNR), polysaccharide deposition method and boiling method in real-time quantitative and provide reference data for clinical lab regarding the PCR kit selection. Methods Three different HBV DNA isolation methods including MNR, polysaccharide deposition method and boiling method were used to study the extraction efficiency and anti-interference ability when HBV DNA virus in lipemia, haemolysis or jaundice serum samples was detected with real-time PCR. The traditional HBV DNA isolation method of Hydroxybenzene-Chloroform was used as control method. Results The quantitative results and amplification efficiency differed among different extraction methods. Complete hemolytic sample brought biggest influence, modest hemolytic sample give minimal influence, whereas the lipemia and jaundice did not bring significant influence to detection. The MNR give the best amplification efficiency and quantitativereproducibility. The extraction efficiency of boiling method was lowest. The fluorescent efficiency of real-timePCR from Hydroxybenzene-Chloroform methods was better than other methods, but it suffered from losingHBV DNA. Conclusions Real-time PCR system of HBV DNA isolation by MNR can provide an accurate,highly sensitive, and rapid method to quantify Hepatitis B virus. It is suitable to use in clinical laboratory.
