摘要
目的探讨荧光定量PCR检测乙型肝炎病毒核酸(HBV-DNA)时核酸提取体系中加入样本量的不同和不同反应体系对检测结果的影响。方法选取乙肝检测患者DNA定量结果为低拷贝数(10-103copies/ml)和高拷贝数(105-107copies/ml)样本各10例,分别按2种核酸提取体系及2种反应体系分为4组进行HBV-DNA荧光定量PCR检测。结果 10例低拷贝数样本减少反应体系后只有3例检出有反应,由于反应体系的减小造成结果存在明显差异,P<0.01;即使增加提取核酸时加入的样本量,结果仍然存在差异,P<0.01。10例高拷贝数的样本改变提取核酸时加入的样本量及最终的反应体系,各组的算术平均值在0.43-2.31倍之间,数量级变化<1。结论反应体系过小会造成低拷贝数样本出现假阴性,所以要保证反应体系中加入足够的提取出的核酸以确保样本HBV-DNA的检出率。
Objective To investigate the influence on HBV-DNA detection results caused by the change of the reac- tion system of disposed samples and the volume of disposed samples in case nucleic acid was extracted. Methods To execute HBV-DNA detection on 10 cases of lower results（10-103copies/ml） and 10 cases of higher results （10s- 10v copies/ml）,which were divided into four groups according to two nucleic acid extraction systems and two reaction system by using FQ-PCR method. Results Only 3 among the total 10 reactive cases of lower result samples were detec- ted when the reaction systems was reduced,which reached the level of significant difference （P^0.01）. Even though the volume of disposed samples were increased,the results still had difference （P^0.01）. When the nucleic acid ex- traction system and the reaction system of higher result samples were changed, the average value of every group was changed from 0.43 times to 2.3i times,and the variation of the results was fallen into one order of magnitude. Conclu- sion Since the smaller reaction system could ~esult in the false negative,we should guarantee that enough nucleic acid was extracted and mixed in reaction system to improve the sample detection rate.
出处
《中国实验诊断学》
2013年第5期877-879,共3页
Chinese Journal of Laboratory Diagnosis
