CN111721945B - 一种用于内分泌室间质评的质控品 - Google Patents
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Abstract
本发明提供一种用于内分泌室间质评的质控品,所述的质控品包括来源于人的血清混合物;所述血清混合物为以下16项内分泌指标中的一项或多项处于高值状态的血清的混合物:FT3、T3、FT4、T4、TSH、VB12、FolateBA、E2‑6III、PRGE孕酮、TSTO、FSH、LH2、PRL催乳素、COR、CPS、IRI。本发明的质控品选择人血清作为基质,在一定程度上减小了质控品与样品间的差异。本发明提供的质控品,制备方法简单,且均匀度和稳定性好,质控偏差小,有较好的应用前景。
Description
技术领域
本发明属于生物检测技术领域,具体涉及一种用于内分泌室间质评的质控品。
背景技术
实验室数据的可靠性是疾病诊断或实验研究有效性的基础,做好实验室的质量控制对检验工作起着重要的作用。室间质量评价(external quality assessment,室间质评)是为客观比较某一实验室测定结果与靶值的差异,由外单位机构采取一定的方法,连续、客观地评价实验室测定的结果,发现误差并校正结果,使各实验室之间的结果具有可比性,是对实验室结果和实验方法的回顾性评价。在室间质评过程中,选择合适的质控品直接影响评价结果。
内分泌项目在临床检测中具有重要意义。甲功类项目有总T3、总T4、促甲状腺激素、游离T3、游离T4等。甲状腺的生理功能主要是促进三大营养物质代谢,调节生长发育,提高组织的耗氧量,促进能量代谢,增加产热和提高基础代谢。当甲状腺功能紊乱时,会发生甲亢或甲减。T3和T4是反应甲状腺功能的主要指标,TSH是反应甲状腺功能状态的指标。
贫血类项目有叶酸、维生素B12等。叶酸是人体内重要的维生素,对蛋白质的合成、DNA、RNA的产生及红细胞、白细胞的合成均起着重要作用,叶酸浓度与胎儿神经管缺陷的发生关系密切。临床检测血清叶酸、维生素B12的水平,结合其他的检测指标,对贫血类型、肝硬化的发生、发展及疗效、预后具有重要的临床价值。
激素类项目有催乳素、雌二醇、孕酮、促卵泡激素、睾酮、促黄体激素、皮质醇等。性激素六项检测是通过测定性激素水平来了解内分泌功能和诊断与内分泌失调相关的疾病。皮质醇在调节脂肪、蛋白质的代谢;免疫系统和其他内分泌系统发挥非常重要的作用,在当前临床医学和药物学的研究中,测定皮质醇的水平是一项重要的指标。
糖尿病相关项目包括:胰岛素、C-肽。胰岛素抗体对糖尿病和低血糖的诊断、鉴别诊断及治疗具有非常重要的意义;测定C-肽浓度,有助于糖尿病的临床分型,了解患者胰岛功能。
由此可见,内分泌对很多疾病的诊断起着非常重要的作用,而做好室内质量控制是保证检测病人结果可靠的前提。长期以来,国内各医疗机构多采用国外内分泌质控品,价格昂贵,对于质控成本的控制有很大的影响。国外如罗氏公司研发的cfas校准品可以为30多项诊断试剂提供校准,为冻干粉状态,稳定性好,包含生化类项目较多,但其应用成本较高。
因此,目前多项内分泌室间质评中,急需一种易获得,成本低,包含内分泌项目多的质控品。
发明内容
为了解决上述问题,本发明提供了一种用于体外诊断的免疫质控品及其制备方法。本发明提供的免疫质控品可以同时检测多种分泌指标,获得方法简单,解决了现有技术中成本高,操作复杂的问题,能够一次性解决多种分泌指标的质评鉴定问题。
一方面,本发明提供了一种用于体外诊断的免疫质控品。
具体地,所述的免疫质控品中包括内分泌指标16项:FT3(游离三碘甲状腺原氨酸)、T3(三碘甲状腺原氨酸)、FT4(游离甲状腺原氨酸)、T4(总甲状腺素)、TSH(促甲状腺素)、VB12(维生素B12)、FolateBA(叶酸)、E2-6III(雌二醇)、PRGE孕酮、TSTO(睾酮)、FSH(卵泡刺激素)、LH2(黄体生成素)、PRL催乳素、COR(皮质醇)、CPS(C肽)、IRI(胰岛素)中的一种或多种。
具体地,所述的质控品中:
FT3含量为:2.8-7.1pmol/L;
T3含量为1.3-3.1nmol/L;
FT4含量为12-22pmol/L;
T4含量为66-181nmol/L;
TSH含量为0.27-4.2mU/L;
VB12含量为180-914pmol/L;
FolateBA含量为11-54nmol/L;
E2-6III含量为7.63-42.6pg/mL;
PRGE孕酮含量为0.7-4.3ng/mL;
TSTO含量为0.9-1.0ng/mL;
FSH含量为1.5-12.4mIU/mL;
LH2含量为1.7-8.6mIU/mL;
PRL含量为4.04-15.2nmol/L;
COR含量为64-536nmol/L;
CPS含量为370-1470nmol/L;
IRI含量为2.6-24.9pmol/L。
具体地,所述的免疫质控品的来源为人血血清。
具体地,所述的免疫质控品中还包括但不限于:防腐剂、稳定剂、抗菌剂、澄清剂,冻干保护剂。
具体地,所述的防腐剂包括但不限于叠氮钠、甲基-异噻唑啉;进一步地,所述的叠氮钠终浓度为0.01%-0.05%。
具体地,所述的稳定剂包括但不限于:PPCD、CCD、SHe-50、AeP-HBC;所述的PPCD的终浓度为2%-4%。
另一方面,本发明提供了一种前述免疫质控品的制备方法,包括以下步骤:
(1)取志愿者血液样本:志愿者血液中内分泌指标的一项或多项处于高值状态;
(2)血清分离:低速离心分离血清备用;
(3)内分泌项目检测:对获得的分离血清进行内分泌项目检测确定各样本中分析项目的值;
(4)血清混合:将步骤血清根据检测值按一定比例混合,使血清中各内分泌指标项达到质控品标准;
(5)添加防腐剂、稳定剂、抗菌剂或澄清剂中的一种或多种。
具体地,所述的步骤(1)中内分泌指标包括FT3(游离三碘甲状腺原氨酸)、T3(三碘甲状腺原氨酸)、FT4(游离甲状腺原氨酸)、T4(总甲状腺素)、TSH(促甲状腺素)、VB12(维生素B12)、FolateBA(叶酸)、E2-6III(雌二醇)、PRGE孕酮、TSTO(睾酮)、FSH(卵泡刺激素)、LH2(黄体生成素)、PRL催乳素、COR(皮质醇)、CPS(C肽)、IRI(胰岛素)中的一种或多种。
具体地,所述的步骤(1)中要求血液材料中人类免疫缺陷病毒(HIV-1和HIV-2)抗体、丙型肝炎病毒HCV抗体、乙型肝炎病毒表面抗原HBsAg、梅毒等检测结果均为阴性。
具体地,所述的步骤(2)低速离心的条件为1000-3000rpm,10-20min;优选为1000rpm,10min。
具体地,所述的步骤(5)中的防腐剂包括但不限于叠氮钠、甲基-异噻唑啉;进一步地,所述的叠氮钠浓度在浓度在0.01%-0.05%;所述的稳定剂包括但不限于:PPCD、CCD、SHe-50、AeP-HBC;所述的PPCD的添加量为终浓度2%-4%。
可选择地,所述的步骤(5)后包括冻干步骤:向步骤(5)得到的产物中添加冻干保护剂后制备成冻干粉;所述的冻干保护剂包括但不限于乙烯乙二醇、甘露醇、甘油、DMSO,DMF;所述的甘露醇的添加终浓度为2%-4%。
在一些实施例中,所述的制备方法包括以下步骤:
(1)取志愿者血液样本:志愿者血液中内分泌指标FT3、T3、FT4、T4、TSH、VB12、FolateBA、E2-6III、PRGE孕酮、TSTO、FSH、LH2、PRL催乳素、COR、CPS、IRI的一项或多项处于高值状态。
(2)血清分离:1000rpm,10min低速离心分离以上样品血清备用;
(3)内分泌项目检测:对获得的分离血清进行内分泌项目检测确定各样本中各分析项目的值。
(4)血清混合:将步骤(1)血清根据检测值按一定比例混合,使血清中各内分泌指标项达到检测标准。
(5)添加终浓度为0.01%的叠氮钠、抗菌剂、澄清剂。
(6)添加终浓度3%的甘露醇作为冻干保护剂后制备冻干粉。
本发明的质控品选择人血清作为基质,在一定程度上减小了质控品与样品间的差异。多种内分泌项目的结合使该质控品可应用于多项检测的室间质量评价中,减少了由于需要的质控品种类较多带来的成本问题。本发明提供的质控品,制备方法简单,且均匀度和稳定性好,质控偏差小,应用价值大。
具体实施方式
下面结合具体实施例,对本发明作进一步详细的阐述,下述实施例不用于限制本发明,仅用于说明本发明。以下实施例中所使用的实验方法如无特殊说明,实施例中未注明具体条件的实验方法,通常按照常规条件,下述实施例中所使用的材料、试剂等,如无特殊说明,均可从商业途径得到。
实施例1一种免疫质控品的制备方法
(1)取志愿者血液样本:志愿者血液中内分泌指标FT3、T3、FT4、T4、TSH、VB12、FolateBA、E2-6III、PRGE孕酮、TSTO、FSH、LH2、PRL催乳素、COR、CPS、IRI的一项或多项处于高值状态。本实施例中各样品信息如下:
(2)血清分离:1000rpm,10min低速离心分离以上样品血清备用;
(3)内分泌项目检测:对获得的分离血清进行内分泌项目检测确定各样本中各分析项目的值。
(4)血清混合:将步骤血清按一定比例混合,使血清中各内分泌指标项达到检测标准。
各样本血清具体添加量如下,总量为1000mL:
样品编号 | 添加量(mL) |
1 | 33 |
2 | 50 |
3 | 39 |
4 | 62 |
5 | 49 |
6 | 20 |
7 | 80 |
8 | 12 |
9 | 26 |
10 | 10 |
11 | 13 |
12 | 86 |
13 | 220 |
14 | 165 |
15 | 135 |
(5)添加终浓度为0.01%的叠氮钠,添加终浓度为2.5%的PPCD,添加终浓度为2.5%的甘露醇。
(6)最终产品中各项内分泌指标数据如下:
实施例2均匀性(复溶稳定性)
对实施例1得到的免疫质控品进行均匀性评价,具体步骤如下:
(1)将质控品加入样本杯中,-20℃冷冻24h。
(2)解冻,在混匀与未混匀两种状态下,对每个项目连续测定5次,计算平均值和标准差。
结果如下表所示:
结果表明:未混匀状态下的均匀性相比混匀状态下的均匀性无明显差异,即在使用过程中可一定程度上降低因操作原因带来的人为误差,更好保证实验结果的可靠性。
实施例3长期稳定性
对实施例1得到的免疫质控品进行长期稳定性评价
将3个平行免疫质控品1-3各置于2-8℃条件下1个月后分别进行测定,并与新制备的质控品进行对比,每个样品每个项目连续测定5次。
数据结果对比如下:
表明本申请提供的血清质控品有良好的稳定性,便于保存,保质期较长。
Claims (4)
1.一种用于内分泌室间质评的质控品,其特征在于,所述的质控品包括来源于人的血清混合物;所述血清混合物为以下16项内分泌指标处于高值状态的血清的混合物:FT3、T3、FT4、T4、TSH、VB12、FolateBA、E2-6III、PRGE孕酮、TSTO、FSH、LH2、PRL催乳素、COR、CPS、IRI;
所述的质控品中
FT3含量为2.8-7.1pmol/L;
T3含量为1.3-3.1nmol/L;
FT4含量为12-22pmol/L;
T4含量为66-181nmol/L;
TSH含量为0.27-4.2mU/L;
VB12含量为180-914pmol/L;
FolateBA含量为11-54nmol/L;
E2-6III含量为7.63-42.6pg/mL;
PRGE孕酮含量为0.7-4.3ng/mL;
TSTO含量为0.9-1.0ng/mL;
FSH含量为1.5-12.4mIU/mL;
LH2含量为1.7-8.6mIU/mL;
PRL含量为4.04-15.2nmol/L;
COR含量为64-536nmol/L;
CPS含量为370-1470nmol/L;
IRI含量为2.6-24.9pmol/L;
所述的质控品还包括防腐剂、稳定剂;
所述的防腐剂为叠氮钠;所述的叠氮钠终浓度为0.01%-0.05%;
所述的稳定剂为PPCD;所述的PPCD的终浓度为2%-4%;
所述的质控品还包括冻干保护剂,所述的冻干保护剂为终浓度2%-4%的甘露醇。
2.一种权利要求1所述质控品的制备方法,其特征在于,包括以下步骤:
(1)取志愿者血液样本:志愿者血液中内分泌指标的一项或多项处于高值状态;
(2)血清分离:低速离心分离血清备用;
(3)内分泌项目检测:对获得的分离血清进行内分泌项目检测确定各样本中分析项目的值;
(4)血清混合:将上述步骤血清根据检测值按一定比例混合,使血清中各内分泌指标项达到权利要求1中的质控品标准;
(5)添加防腐剂、稳定剂;
(6)添加终浓度2%-4%的甘露醇作为冻干保护剂后制备成冻干粉。
3.根据权利要求2所述的制备方法,其特征在于,所述的步骤(1)中内分泌指标为FT3、T3、FT4、T4、TSH、VB12、FolateBA、E2-6III、PRGE孕酮、TSTO、FSH、LH2、PRL催乳素、COR、CPS、IRI;所述的步骤(2)低速离心的条件为1000-3000 rpm,10-20min。
4.权利要求1所述的质控品和/或权利要求2-3中任一项所述的制备方法在内分泌室间质评中的应用。
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