高载量标本HBV-DNA定量检测时稀释液的选择与评价  

Select and evaluate the diluents for quantitative detection of HBV DNA of high loads sample

作　　者：汪东剑[1] 余维涛[1] 张晓云[1] 刘冬萍[1] 邱华琴[1] 彭慧琼[1] 

机构地区：[1]中国人民解放军第一八四医院输血科,鹰潭335000

出　　处：《中华实验和临床病毒学杂志》2016年第2期223-227,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：南京军区医药卫生科研基金课题（M5078）

摘　　要：目的 对高载量标本HBV-DNA稀释检测时稀释液进行选择和评价,期望筛选出较为理想适用于临床的稀释液.方法 以阴性质控品作为标准稀释液,阴性血清、生理盐水、蒸馏水作为候选稀释液,分别对定值标准品（浓度为2.00×10^9 IU/ml）进行10倍、100倍的稀释检测,比较使用候选稀释液与标准稀释液的检测结果,同时进行偏差分析.结果 在10倍稀释时,使用蒸馏水稀释与标准稀释液稀释,其检测结果间差异无统计学意义（=2.04,P＞0.05）,偏差小于行业标准规定的总允许误差;使用阴性血清、蒸馏水时检测结果高于标准稀释（P＜0.05）,分别有16.67％和20.00％的偏差超过行业标准规定的总允许误差.在100倍稀释时,使用待选稀释液,其检测结果均高于使用标准稀释液时,差异有统计学意义（P＜0.05）,且以蒸馏水作为稀释液时的检测结果与以标准稀释液稀释时的检测结果呈良好的线性关系,其公式为Y=0.963X＋0.267;使用不同稀释液时偏差均小于行业标准规定的总允许误差,以标准稀释液偏差最小,其次为蒸馏水、生理盐水、阴性血清（P＜0.05）.结论 在10倍稀释时,最适稀释液为阴性质控品与蒸馏水;在100倍稀释时,最适稀释液为阴性质控品,其次为蒸馏水、生理盐水、阴性血清,以蒸馏水作为稀释液时,可通过公式y=0.963X＋0.267对检测结果进行校正,确保结果更为精确.Objective To select and evaluate the diluents for quantitative detection of HBV DNA of high loads sample,hope to find the most applicable diluents which could be used in clinical test.Methods The standard substance（2.00× 109IU/ml）was 10 and 100 times diluted by different diluents,compare the result of test,and the bias was analysis taking negative quality control as standard diluents,negative serum,physiological saline,and distilled water as candidate diluents.Results When 10 times diluted,there was no statistically difference between the standard diluents and distilled water as diluents（t =2.04,P 〉 0.05）,the bias were less than the TEa regulated by professional standard.When used negative serum and physiological saline as diluents,the results were higher than that of standard diluents （P 〈 0.05）,and the ratio of the bias higher than the TEa was 16.67% and 20.00%.When 100 times diluted,the results of candidate diluents were higher than that of standard diluents.In this time,the result of distilled water diluted detection presented a good linear relationship with the result of standard diluted detection,the formula was Y =0.963X ＋ 0.267 （Y =result of standard diluted detection,X =result of distilled water diluted detection）.All the bias were less than the TEa,the sequence of bias sort by ascending counts were negative quality control,distilled water,physiological saline and negative serum.Conclusions The most applicable diluents were negative quality control and distilled water with 10 times dilution.When 100 times diluted was used,the most applicable diluents was negative quality control,then was distilled water,physiological saline and negative serum.If using the distilled water to dilute,we could corrected the result by the formula Y =0.963X ＋ 0.267 to ensure the result to be more exactly.

关 键 词：稀释液 乙型肝炎病毒 DNA 检测 

分 类 号：R512.62[医药卫生—内科学]