HCV核糖核酸国家二级标准物质的研制

Preparation for national secondary reference material of HCV RNA

目的研制HCV RNA国家二级标准物质。方法用HCV阴性血清将阳性血清稀释至浓度约3.0×10^5IU/m L,按每支1 m L分装,真空冷冻干燥。制备后进行均匀性和稳定性检验,并做临床适用性评价。定值溯源至国际标准物质（编号06/102）。结果均匀性结果显示瓶间不精密度1.77%,瓶内不精密度1.40%（F=1.596 9,P〉0.05）;20～25℃稳定14 d,2～8℃稳定2个月,-20℃稳定12个月（P〉0.05）;37℃不稳定（P〈0.05）。开瓶后放置7 d、反复冻融5次和模拟运输后检测值与对照组（-20℃保存）比较,差异无统计学意义（P〉0.05）。定值为（1.9±0.9）×10^5IU/m L。结论制备物达到了国家二级标准物质的要求,可以作为核酸扩增检测的HCV RNA标准物质。

Objective To prepare the national secondary reference material of HCV RNA for nucleic acid amplification technology.Methods The candidate sample of HCV RNA-positive plasma was diluted to concentration of approximately 3. 0 × 10^5 IU / ml with HCV-negative human plasma and then lyophilized and dispensed as 1 m L for each tube. The homogeneity and stability of the prepared reference materials were tested and the clinical applicability was evaluated. The quantity of prepared material was traced to the international standard（ 06 /102）. Results The evaluation of homogeneity indicated the between-run precision was 1. 77% and the within-run precision was 1. 40%（ F = 1. 596 9,P〉0. 05）. The prepared materials could be stable at room temperature（ 20-25 ℃） for 14 days,at 2-8 ℃ for 2 months and at-20 ℃ for 12 months（ P〉0. 05）,but was unstable at 37 ℃（ P〈0. 05）. Under the conditions of opening the tube after 1 week,repeatedly thawing / freezing for 5 cycles and simulated transportation between two laboratories,no significant differences were observed in the comparisons of the test results of the prepared materials with controls stored at- 20 ℃（ P〉0. 05）.The quantity of the prepared material of HCV RNA was determined as（ 1. 9 ± 0. 9） × 10^5 IU / m L. Conclusion The prepared material of HCV RNA reached the requirements of the national secondary reference and can be used as the standard for nucleic acid amplification test of HCV RNA.