CAJ | 학술논문

目的：探讨浓缩液剂量、沉淀混匀方式和裂解煮沸时间等前处理因素对实时荧光定量 PCR法检测乙型肝炎病毒DNA（HBV‐DNA）的影响。方法在浓缩液分别为90、95、105、110μL ；混匀方式分别为：漩涡混匀15 s组和30 s组，56℃预热120 s再漩涡混匀15 s组和56℃预热180 s再漩涡混匀15 s组；裂解煮沸时间分别为：6，8，12，14 min等条件下对80例自配的低值质控品，浓度在103～104 IU／mL进行测定，并与标准操作即浓缩液100μL ，镊子敲打混匀15 s ，裂解煮沸时间10 min比较。结果漩涡混匀15 s组和56℃预热180 s再漩涡混匀15 s与标准组比较差异有统计学意义（P＜0．05）；煮沸裂解时间6 min组与10 min组间比较差异有统计学意义（P＜0．05）。结论在一定范围内适当改变浓缩液剂量、混匀方式和煮沸裂解时间，不会影响检测结果。但经过一定时间预热处理后再振荡混匀的方式更容易使沉淀混匀和核酸的释放。
목적：탐토농축액제량、침정혼균방식화렬해자비시간등전처리인소대실시형광정량 PCR법검측을형간염병독DNA（HBV‐DNA）적영향。방법재농축액분별위90、95、105、110μL ；혼균방식분별위：선와혼균15 s조화30 s조，56℃예열120 s재선와혼균15 s조화56℃예열180 s재선와혼균15 s조；렬해자비시간분별위：6，8，12，14 min등조건하대80례자배적저치질공품，농도재103～104 IU／mL진행측정，병여표준조작즉농축액100μL ，섭자고타혼균15 s ，렬해자비시간10 min비교。결과선와혼균15 s조화56℃예열180 s재선와혼균15 s여표준조비교차이유통계학의의（P＜0．05）；자비렬해시간6 min조여10 min조간비교차이유통계학의의（P＜0．05）。결론재일정범위내괄당개변농축액제량、혼균방식화자비렬해시간，불회영향검측결과。단경과일정시간예열처리후재진탕혼균적방식경용역사침정혼균화핵산적석방。
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 .