人脲原体核酸检测试剂盒国家参考品的制备

发布时间：2018-03-16 16:01

  本文选题：脲原体　切入点：血清型　出处：《中国生物制品学杂志》2014年08期 　论文类型：期刊论文

【摘要】：目的制备人脲原体核酸检测试剂盒国家参考品。方法培养人脲原体两大生物群14个标准血清型菌株,培养液灭活后经颜色改变单位(color change unit,CCU)法与PCR相结合的方法进行定值,组成能覆盖脲原体所有表型及基因型的14份阳性参考品P1-P14(浓度为106 CCU/ml)及14份检测限参考品L1-L14(浓度为104 CCU/ml);选择6份能排除脲原体核酸检测试剂盒非特异性或交叉反应的样品组成阴性参考品N1-N6;选择生物群1的微小脲原体(ureaplasma parvum,UP)1和UP14作为重复性参考品R1、R2(浓度分别为105和104 CCU/ml)。用5家公司生产的脲原体核酸检测试剂盒对参考品进行验证,确定准确性、特异性、检测限、重复性、稀释线性的性能指标;并考察参考品在不同条件下(2~8℃放置7 d,37℃放置3、7、12 d,-20℃及常温反复冻融5次)的稳定性。结果支原体各培养液经CCU法测定,浓度均在106~107 CCU/ml之间,以CCU法测定浓度为靶值,绝对偏差均在±0.5个数量级以内。4家公司生产的不同试剂盒检测参考品,在准确性、特异性、检测限及重复性上均符合要求;在稀释线性上,最低稀释浓度为102 CCU/ml的样品只有2家可以检出,其他几家的稀释线性相关系数以103~106 4个浓度计算,r值均0.990 0;1家(RNA检测)检测R2的重复性,CV为5.5%。经不同条件处理的参考品的稳定性有轻微变化,但均处于设定的可接受范围内。结论制备了人脲原体核酸检测试剂盒国家参考品,该参考品可满足人脲原体核酸分型、定性及定量检测要求,经多家实验室进行验证,可用于国内大多数试剂盒的性能评价及临床实验室质量评价。
[Abstract]:Objective to prepare the national reference kit for detection of human Ureaplasma Urealyticum nucleic acid. Methods 14 standard serotypes of two biota of Ureaplasma Urealyticum were cultured. The culture medium was inactivated and determined by the method of color change unit (change) and PCR. 14 positive reference materials covering all phenotypes and genotypes of Ureaplasma Urealyticum P1-P14 (concentration 106 CCU / ml) and 14 limited reference materials L1-L14 (concentration 104 CCU / ml); 6 samples selected to exclude nonspecific or cross reaction of Ureaplasma Urealyticum nucleic acid detection kit. The samples were composed of negative reference material N1-N6; ureaplasma parvum UP1 and UP14 from biome 1 were selected as repeatable reference materials R1UP R2 (concentrations 105 and 104CCU / ml, respectively. The reference was verified with Ureaplasma Urealyticum nucleic acid detection kit produced by five companies, respectively. Determine the accuracy, specificity, detection limit, repeatability, dilution linear performance indicators; The stability of the reference was investigated under different conditions at 8 鈩，

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