Explore the influence factors of hepatitis B virus DNA quantitative testing in nucleic acid extraction process
10.3969/j.issn.1673-4130.2015.15.013
- VernacularTitle:HBV-DNA 定量检测核酸提取过程中影响因素的探讨
- Author:
Ying PENG
;
Xianyong WEN
;
Zhenghua DENG
- Publication Type:Journal Article
- Keywords:
polymerase chain reaction;
concentrate dose;
the time of oscillation;
boiling time
- From:
International Journal of Laboratory Medicine
2015;(15):2156-2157
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the concentrated liquid doses ,oscillation intensity and cracking boiling time to the influence of pre‐treatment HBV‐DNA .Methods The concentrate doses was 90 ,95 ,105 ,110 μL respectively ;mixing methods :vortex mixing 15 seconds group and 30 seconds group ,56 ℃ preheated 120 seconds and then vortex mix 15 seconds group ,56 ℃ preheated 180 seconds and then swirl mix 15 seconds group ;cracking boiling time was 6 ,8 min ,under 12 min ,14 min respectively .Under the a‐bove conditions ,80 samples prepared by the laboratory were measured ,comparing with standard operating ,concentrate 100μL ,beat mix 15 seconds ,cracking boiling 10 min ,which HBV‐DNA concentration was 103 -104 IU/mL .Results Vortex mixing 15 s group and 56 ℃ preheat 180 s then swirl mixing 15 s were both a significant difference(P<0 .05) in the mix mode groups ,comparing with the standard operating group .There were significant differences between the groups boiling lysis time of 6 min and 10 min group ( P<0 .05) .Conclusion Within a certain range ,changing the dose of the concentrated solution ,mixing mode and the time of boiling lysis in extraction process of HBV‐DNA does not affect the test results ,But mixing ways after a certain period preheating make it easier to mix precipitation and release nucleic acid .
