CAJ | 학술논문

实时PCR技术因其快速、准确、灵敏度和重复性高、可减少交叉污染等特点而广泛应用于分子生物学和医学研究领域。本研究建立了一种基于LUX （Light Upon eXtension）引物的HBV病毒载量检测的实时定量PCR检测方法。通过检测系列稀释的HBV DNA（5-5×108拷贝/反应）来验证LUX实时分析的性能和灵敏度。结果表明该检测方法在Ct值和log10 HBV DNA浓度之间存在很好的线形关系，并且具有很高的灵敏度，检测低限可达每毫升血清中50拷贝的HBV。对91份阳性血清样品的检测和熔解曲线分析表明该方法具有很高的特异性。新建立的LUX实时检测方法为检测治疗效果、研究HBV病毒载量和疾病发展之间的关系提供了一种理想的工具。
실시PCR기술인기쾌속、준학、령민도화중복성고、가감소교차오염등특점이엄범응용우분자생물학화의학연구영역。본연구건립료일충기우LUX （Light Upon eXtension）인물적HBV병독재량검측적실시정량PCR검측방법。통과검측계렬희석적HBV DNA（5-5×108고패/반응）래험증LUX실시분석적성능화령민도。결과표명해검측방법재Ct치화log10 HBV DNA농도지간존재흔호적선형관계，병차구유흔고적령민도，검측저한가체매호승혈청중50고패적HBV。대91빈양성혈청양품적검측화용해곡선분석표명해방법구유흔고적특이성。신건립적LUX실시검측방법위검측치료효과、연구HBV병독재량화질병발전지간적관계제공료일충이상적공구。
Real-time PCR technology has been widely used in molecular biology and medical research due to its im-proved rapidity,accuracy,sensitivity,reproducibility and reduced risk of carry-over contamination.A quantitative real-time PCR method based on Light Upon Extension (LUX)primer was established to measure the viral load of hepatitis B virus in serum.The performance of the LUX real-time assay was validated by testing serial dilutions of HBV DNA (5 to 5 ×108 copies per reaction)and a good linear relationship was obtained between the Ct values and the log10 concentration of the HBV DNA.The LUX method possessed high sensitivity and the detection limit of this system was as few as 25 copies/mL of serum.9 1 positive serum samples were detected to further evaluate the assay and the high specificity was confirmed by melting curve analysis.This assay provides an ideal approach for monitoring the treatment efficacy and studying the relationship between HBV viral load and stage of disease.