快速血浆反应素环状卡片试验实验室内质控品的制备及稳定性评价
|
摘要
目的制备快速血浆反应素环状卡片试验(rapid plasma reagin circle card test,RPR)实验室内质控品,并对其稳定性进行评价。方法选择肝炎病毒标志物、梅毒螺旋体特异性抗体/非特异性抗体和HIV抗体均为阴性的血清,选择肝炎病毒标志物和HIV抗体均为阴性,梅毒螺旋体非特异性抗体为阳性的血清,将阳性和阴性血清分别混匀,2 643×g离心10 min去除蛋白质沉淀,56℃加热30 min灭活传染性病原体,经0. 2μm滤膜过滤除菌。阴性质控品使用RPR阴性血清基质制备;用阴性血清基质稀释阳性血清基质制备1∶2~+阳性质控品。将阴性和阳性质控品按7 d使用量分装,冷冻保存在-70和-20℃,检测6个月冷冻保存的稳定性及2～8℃保存1周的开瓶稳定性,并进行瓶间差验证。结果质控品-70和-20℃6个月冷冻保存的稳定性及复融后2～8℃保存1周的开瓶稳定性均良好;两种储存条件保存的质控品均无甁间差。结论自制的RPR试验室内质控品6个月内稳定性和均一性均良好,能满足临床实验室日常质控活动。
Objective To prepare the internal quality controls for rapid plasma regain circle card test(RPR)and evaluate their stability. Methods The serum samples negative for hepatitis virus markers,specific/non-specific Treponema pallidum antibodies and HIV antibody,as well as those negative for hepatitis virus markers and HIV antibody but positive for non-specific T. pallidum antibody were pooled separately,centrifuged at 2 643 × g for 10 min to remove the protein precipitates,heated at 56 ℃ for 30 min to inactivate the infectious pathogens,and sterilized by filtration with a 0. 2 μm membrane. Negative quality controls were prepared with the serum matrix negative in RPR test,while the positive serum matrix was diluted with negative serum matrix to obtain 1 ∶ 2~+positive controls. The negative and positive quality controls were filled according to the volumes used in 7 d,and stored at-70 ℃ and-20 ℃ in frozen. The stabilities after storage in frozen for 6 months and after storage at 2 ～ 8 ℃ for one week were evaluated,of which the difference between containers was verified. Results The quality controls showed high stability after storage at-70 ℃ and-20 ℃ in frozen for 6 months and after storage at 2 ～ 8 ℃ for one week,which showed no significant difference between containers.Conclusion The prepared internal quality controls showed high stability and homogeneity within 6 months,which met the requirements for daily quality control in clinical laboratory.
Objective To prepare the internal quality controls for rapid plasma regain circle card test(RPR)and evaluate their stability. Methods The serum samples negative for hepatitis virus markers,specific/non-specific Treponema pallidum antibodies and HIV antibody,as well as those negative for hepatitis virus markers and HIV antibody but positive for non-specific T. pallidum antibody were pooled separately,centrifuged at 2 643 × g for 10 min to remove the protein precipitates,heated at 56 ℃ for 30 min to inactivate the infectious pathogens,and sterilized by filtration with a 0. 2 μm membrane. Negative quality controls were prepared with the serum matrix negative in RPR test,while the positive serum matrix was diluted with negative serum matrix to obtain 1 ∶ 2~+positive controls. The negative and positive quality controls were filled according to the volumes used in 7 d,and stored at-70 ℃ and-20 ℃ in frozen. The stabilities after storage in frozen for 6 months and after storage at 2 ～ 8 ℃ for one week were evaluated,of which the difference between containers was verified. Results The quality controls showed high stability after storage at-70 ℃ and-20 ℃ in frozen for 6 months and after storage at 2 ～ 8 ℃ for one week,which showed no significant difference between containers.Conclusion The prepared internal quality controls showed high stability and homogeneity within 6 months,which met the requirements for daily quality control in clinical laboratory.
引文
[1] YE Y W,WANG Y S,SHEN Z Y. National Guide to Clinical Laboratory Procedures 3rd ed.[M]. Nanjing:Southeast University Press,2006:647-650.(in Chinese)叶应妩,王毓三,申子瑜.全国临床检验操作规程[M]. 3版.江苏南京:东南大学出版社,2006:647-650.
[2] China National Accreditation Service for Conformity Assessment(CNAS). Accreditation Criteria for the Quality and Competence of Medical Laboratories(ISO 15189:2010,IDT)(3rd)[S/OL].(2013-11-22)[2018-04-02]. https://www.cnas.org.cn/rkgf/sysrk/jbzz/2013/12/750592.shtml.2012.(in Chinese)中国合格评定国家认可委员会.医学实验室质量和能力认可准则(ISO 15189:2012,IDT)(第三版)[S/OL]. https://www.cnas.org.cn/rkgf/sysrk/jbzz/2013/12/750592.shtml. 2012.
[3] China National Accreditation Service for Conformity Assessment(CNAS). Guidance on the Application of Accreditation Criteria for the Medical Laboratory Quality and Competence in the Field of Clinical Qualitative Immunology[S/OL].(2018-03-01)[2018-04-02]. https://www.cnas.org.cn/images/rkgf/sysrk/rkyyzz/2018/03/01/B33603-67CC1BFC351B14EB225A246298.pdf.CNAS-CL02-A004,2018.中国合格评定国家认可委员会.医学实验室质量和能力认可准则在临床免疫学定性检验领域的应用说明[S].(2018-03-01)[2018-04-02]. https://www.cnas.org.cn/images/rkgf/sysrk/rkyyzz/2018/03/01/B3360367CC1BFC351B14EB225A246298.pdf.CNAS-CL02-A004.2018.
[4]潘敏,沈菁,李山,等.定值多项目复合免疫质控品制备的研究[J].广西医科大学学报,2015,32(5):756-758.
[5] RIXIN JJAMTSHO R. Stability of lyophilized human serum for use as quality control material in Bhutan[J]. Indian J Clin Biochem,2013,28(4):418-421.
[6] ZHANG M,LU H X,XIN Q,et al. Preparation and evaluation of laboratory internal quality control materials for screening of irregular antibody[J]. Chin J Clin Lab Sci,2015,33(12):929-931.(in Chinese)张敏,卢慧霞,辛琪,等.不规则抗体筛查室内质控品的制备及初步应用[J].临床检验杂志,2015,33(12):929-931.
[7] MA C Y,YU Y,GUAN X Z,et al. Evaluation of internal quality control products for blood transfusion compatibity testing[J].Chin J Blood Transfusion,2013,26(5):483-487.(in Chinese)马春娅,于洋,关晓珍,等.输血相容性检测室内质控品的临床应用评价[J].中国输血杂志,2013,26(5):483-487.
[8]杨凤英,梅霓,高晓丽.乙型肝炎病毒表面抗原室内质控血清制备及评价[J].河南预防医学杂志,2014,25(6):446-447,464.
[9] CHEN B,WANG M Y,PEI L J,et al. The preparation and application of the internal quality control sample in human immunodeficiency virus rapid test[J]. Chin J AIDS STD,2015,21(7):608-610.(in Chinese)陈兵,王美玉,裴丽健,等. HIV抗体快速诊断室内质控品的制备及应用[J].中国艾滋病性病,2015,21(7):608-610.
[10]周志米,张晓燕.自制梅毒、丙肝、艾滋病抗体复合质控品的稳定性评价[J].浙江实用医学,2012,17(4):301-302.
[11] CHEN S B,HE Z Y,YU L,et al. Application and analysis of multiple-label internal quality control in nucleic acid blood screening[J]. Chin J Blood Transfusion,2016,29(4):423-426.(in Chinese)陈少彬,何子毅,余霖,等.多标记室内质控品在血液核酸筛查的应用分析[J].中国输血杂志,2016,29(4):423-426.
[12] LEI X X,MING K H,LIN G Y,et al. Evaluation of the stability of self-preparing internal quality control materials of HCVRNA[J]. J Trop Med,2015,15(3):321-325.(in Chinese)雷秀霞,明凯华,林桂莹,等.自配HCV-RNA室内质控品稳定性探讨[J].热带医学杂志,2015,15(3):321-325.
[13] WANG X L,LIU F,JIANG L L,et al. Preparation and application of intrtnal quality control material for nucleic acid testing of human papillomavirus type 16 and 18[J]. Chin J Clin Lab Sci,2016,34(1):56-59.(in Chinese)王雪亮,刘芬,蒋玲丽,等.人乳头瘤病毒16和18型核酸检测用质控品制备及应用[J].临床检验杂志,2016,34(1):56-59.
[14] WEI Z H,GUO R,HUANG H Y,et al. Preparation and effectiveness evaluation on internal quality control for quantitative determination of hepatitis B virus DNA[J]. Chin J Biologicals,2016,29(2):163-166.(in Chinese)魏振宏,郭瑞,黄焕源,等.乙型肝炎病毒DNA定量检测室内质控品的制备及效果评价[J].中国生物制品学杂志,2016,29(2):163-166.
[2] China National Accreditation Service for Conformity Assessment(CNAS). Accreditation Criteria for the Quality and Competence of Medical Laboratories(ISO 15189:2010,IDT)(3rd)[S/OL].(2013-11-22)[2018-04-02]. https://www.cnas.org.cn/rkgf/sysrk/jbzz/2013/12/750592.shtml.2012.(in Chinese)中国合格评定国家认可委员会.医学实验室质量和能力认可准则(ISO 15189:2012,IDT)(第三版)[S/OL]. https://www.cnas.org.cn/rkgf/sysrk/jbzz/2013/12/750592.shtml. 2012.
[3] China National Accreditation Service for Conformity Assessment(CNAS). Guidance on the Application of Accreditation Criteria for the Medical Laboratory Quality and Competence in the Field of Clinical Qualitative Immunology[S/OL].(2018-03-01)[2018-04-02]. https://www.cnas.org.cn/images/rkgf/sysrk/rkyyzz/2018/03/01/B33603-67CC1BFC351B14EB225A246298.pdf.CNAS-CL02-A004,2018.中国合格评定国家认可委员会.医学实验室质量和能力认可准则在临床免疫学定性检验领域的应用说明[S].(2018-03-01)[2018-04-02]. https://www.cnas.org.cn/images/rkgf/sysrk/rkyyzz/2018/03/01/B3360367CC1BFC351B14EB225A246298.pdf.CNAS-CL02-A004.2018.
[4]潘敏,沈菁,李山,等.定值多项目复合免疫质控品制备的研究[J].广西医科大学学报,2015,32(5):756-758.
[5] RIXIN JJAMTSHO R. Stability of lyophilized human serum for use as quality control material in Bhutan[J]. Indian J Clin Biochem,2013,28(4):418-421.
[6] ZHANG M,LU H X,XIN Q,et al. Preparation and evaluation of laboratory internal quality control materials for screening of irregular antibody[J]. Chin J Clin Lab Sci,2015,33(12):929-931.(in Chinese)张敏,卢慧霞,辛琪,等.不规则抗体筛查室内质控品的制备及初步应用[J].临床检验杂志,2015,33(12):929-931.
[7] MA C Y,YU Y,GUAN X Z,et al. Evaluation of internal quality control products for blood transfusion compatibity testing[J].Chin J Blood Transfusion,2013,26(5):483-487.(in Chinese)马春娅,于洋,关晓珍,等.输血相容性检测室内质控品的临床应用评价[J].中国输血杂志,2013,26(5):483-487.
[8]杨凤英,梅霓,高晓丽.乙型肝炎病毒表面抗原室内质控血清制备及评价[J].河南预防医学杂志,2014,25(6):446-447,464.
[9] CHEN B,WANG M Y,PEI L J,et al. The preparation and application of the internal quality control sample in human immunodeficiency virus rapid test[J]. Chin J AIDS STD,2015,21(7):608-610.(in Chinese)陈兵,王美玉,裴丽健,等. HIV抗体快速诊断室内质控品的制备及应用[J].中国艾滋病性病,2015,21(7):608-610.
[10]周志米,张晓燕.自制梅毒、丙肝、艾滋病抗体复合质控品的稳定性评价[J].浙江实用医学,2012,17(4):301-302.
[11] CHEN S B,HE Z Y,YU L,et al. Application and analysis of multiple-label internal quality control in nucleic acid blood screening[J]. Chin J Blood Transfusion,2016,29(4):423-426.(in Chinese)陈少彬,何子毅,余霖,等.多标记室内质控品在血液核酸筛查的应用分析[J].中国输血杂志,2016,29(4):423-426.
[12] LEI X X,MING K H,LIN G Y,et al. Evaluation of the stability of self-preparing internal quality control materials of HCVRNA[J]. J Trop Med,2015,15(3):321-325.(in Chinese)雷秀霞,明凯华,林桂莹,等.自配HCV-RNA室内质控品稳定性探讨[J].热带医学杂志,2015,15(3):321-325.
[13] WANG X L,LIU F,JIANG L L,et al. Preparation and application of intrtnal quality control material for nucleic acid testing of human papillomavirus type 16 and 18[J]. Chin J Clin Lab Sci,2016,34(1):56-59.(in Chinese)王雪亮,刘芬,蒋玲丽,等.人乳头瘤病毒16和18型核酸检测用质控品制备及应用[J].临床检验杂志,2016,34(1):56-59.
[14] WEI Z H,GUO R,HUANG H Y,et al. Preparation and effectiveness evaluation on internal quality control for quantitative determination of hepatitis B virus DNA[J]. Chin J Biologicals,2016,29(2):163-166.(in Chinese)魏振宏,郭瑞,黄焕源,等.乙型肝炎病毒DNA定量检测室内质控品的制备及效果评价[J].中国生物制品学杂志,2016,29(2):163-166.