实时荧光定量PCR检测巨细胞病毒两种白细胞提取方法对比分析  

The analysis of two methods extracting lymphocyte for the cytomegalovirus detection by fluorescence quantified PCR

作　　者：刘广印[1] 周光庭[1] 沈佐君[2] 

机构地区：[1]安徽省亳州市人民医院中心实验室,236800 [2]安徽省临床检验中心,安徽合肥230001

出　　处：《淮海医药》2007年第2期95-96,共2页Journal of Huaihai Medicine

基　　金：安徽省自然科学基金资助项目(050430902)

摘　　要：目的比较实时荧光定量PCR技术检测巨细胞病毒两种白细胞提取处理方法。方法对51例新生儿黄疸患者外周血分别进行淋巴细胞分离液法提取白细胞和0.8%氯化胺裂解红细胞法提取白细胞,然后同时进行实时荧光定量PCR检测。结果51例样本中两种方法均阳性5例,白细胞抽提液法阳性、氯化胺裂解红细胞法阴性2例;白细胞抽提液法阴性、氯化胺裂解红细胞法阳性4例;两者均阴性40例;氯化胺裂解红细胞阳性率法17.64%,淋巴细胞分离液法阳性率13.72%。经统计学分析,两方法阳性率差异无显著性(χ2=0.5,P>0.05)。结论氯化胺裂解红细胞法提取白细胞方法简便、可靠,可用于荧光定量PCR检测巨细胞病毒。Objective To compare the two methods of extracting lymphocyte in the detecting of cytomegalovirus by real-time fluorescence quantified PCR. Methods Real-time fluorescence quantified PCR detect of cytomeglovirus in lymphocyte was conducted at the same time after peripheral blood samples of 51 neonatal jaundice were separated by lymphocyte apart fluid or 0.8 % NH4Cl. Results Among 51 samples, 5 were detected CMV positive by the two methods;2 were CMV positive by lymphocyte apart fluid but negative by 0.8%NH4Cl;4 were positive by 0.8o/oo NH4Cl but negative by lymphocyte apart fluid ; 40 were positive by two methods. Positive rate of method of 0.8 % NH4Cl was 17.64% compared with 13.72 % of method of lymphocyte apart fluid. The difference was not significant （χ2= 0.5,P〉0. 05）. Conclusion Methods of ripper erythrocyte by 0.8% NH4Cl was simple and reliable. It can be used to detect cytomegalovirus by real-time fluorescence quantified PCR.

关 键 词：实时荧光定量PCR 巨细胞病毒 淋巴细胞分离液法 氯化胺裂解红细胞法 

分 类 号：R373[医药卫生—病原生物学]