CAJ | 학술논문

目的:应用两种不同方法检测系统性红斑狼疮(SLE)患者CD4~+T细胞p16基因甲基化状态,比较两种检测方法的差异.方法:采用以Taqman探针为基础的MSP法(方法1)和以SYBR Green I为基础的MSP法(方法2)分别检测40例SLE患者和20例正常人CD4~+T细胞中p16基因启动子区甲基化状态.结果:Taqman探针方法的结果为:SLE患者CD4~+T细胞p16基因甲基化阳性率(35.7%,10/28)高于对照组(10%,2/20),两者比较差异有统计学意义(χ~2=4.11,P＜0.05).SYBR Green I方法的结果为:患者组和对照组的CD4~+T细胞的p16基因均呈高甲基化状态,应用t检验分析发现P＞0.05,二者无统计学差异.结论:Taqman探针法消除了引物二聚体和非特异性扩增对试验结果的影响,提高了结果的特异性和准确性,被证明是进行DNA甲基化状态检测的可靠方法.
목적:응용량충불동방법검측계통성홍반랑창(SLE)환자CD4~+T세포p16기인갑기화상태,비교량충검측방법적차이.방법:채용이Taqman탐침위기출적MSP법(방법1)화이SYBR Green I위기출적MSP법(방법2)분별검측40례SLE환자화20례정상인CD4~+T세포중p16기인계동자구갑기화상태.결과:Taqman탐침방법적결과위:SLE환자CD4~+T세포p16기인갑기화양성솔(35.7%,10/28)고우대조조(10%,2/20),량자비교차이유통계학의의(χ~2=4.11,P＜0.05).SYBR Green I방법적결과위:환자조화대조조적CD4~+T세포적p16기인균정고갑기화상태,응용t검험분석발현P＞0.05,이자무통계학차이.결론:Taqman탐침법소제료인물이취체화비특이성확증대시험결과적영향,제고료결과적특이성화준학성,피증명시진행DNA갑기화상태검측적가고방법.
Objective:To use two kinds of real-time PCR to detect the methylation status of p16 gene in CD4~+ T cell derived from SLE patients and compare the effect of the two methods. Methotis:P16 promotor methylation in CD4'T cell was detected with both the Taqman probe based real-time PCR technology and the SYBR Green I based real-time PCR technology(method 2)in 40 SLE patients and 20 healthy controls.Results:The result of Taqnmn probe method showed that the rate of p16 gene hypermethylation was higher in SLE pmienm(35.7%)than in that of the controls(10%)(χ~2=4.11,P=0.04＜0.05).The result of SYBR Green method showed that there was no significant difference between the patients and controls(P＞0.05).Conclusions:Taqman probe method can effectively eliminate the effect of the primer-dimers(PDs)and nonspecific amplieation on the process of PCR,and increase the speeificity and accuracy of the result.This method can be used to detect the the methylmion status of DNA.