人绒毛膜促性腺激素第三次免疫测定用国家标准品候选品的建立

目的 建立人绒毛膜促性腺激素(HCG)第三次免疫测定用国家标准品候选品.方法 精确量取来源于妊娠妇女尿液的高纯HCG原料、人血白蛋白和海藻糖,加入至磷酸缓冲液(0.05mol/L,pH =7.4)内,充分混匀后置于2～8℃过夜,次日无菌过滤,0.5mL分装后,冻干制备成HCG国家标准品候选品.以HCG第五次国际标准品(07/364)为对照品,采用9个厂家10种HCG检测试剂盒进行协作标定.同时对候选品的瓶间精密度、稳定性进行分析.结果 HCG候选品免疫活性为14.8±2.5IU/支,瓶间精密度为2.38％,37℃放置16周后,HCG候选品的免疫活性下降在10％范围内.结论 候选品各项指标均符合要求,可作为HCG第三次免疫测定用国家标准品使用.本标准品的建立,将为HCG免疫测定用试剂盒的标准化提供定量基础.     

甲状腺素（T4）第三次免疫测定用国家标准品的建立

目的：建立甲状腺素（T4）第三次免疫测定用国家标准品。方法和结果：精确称取高纯T4原料,用含1．0％BSA的0．02MPBS缓冲液配制成2000ng／ml溶液,经无菌过滤处理后,分装、冻干制成候选品。以T4第二次国家标准品（150551-0004）为对照品,经协作标定,候选品中T4免疫活性为940ng／支。结论：候选品各项指标均符合要求,可作为T4第三次免疫测定用国家标准品使用。本标准品的建立,为T4免疫测定用试剂盒的标准化提供定量基础。     

Anti-TgAb和Anti-TpoAb测定用国家标准品的建立

目的:为规范国产Anti-TgAb 和Anti-TpoAb定量测定试剂盒,本实验室研制并建立了Anti-TgAb 和Anti-TpoAb免疫测定用国家标准品.方法和结果:选择经临床检测Anti-TgAb、Anti-TpoAb高值血清,经离心、过滤等处理后,分装、冻干制成标准品候选品.以WHO Anti-TgAb 1st IRP (65/093)为对照品,经协作标定,候选品中Anti-TgAb免疫活性均值为3691IU/支,以WHO Anti-TpoAb RS (66/387)为对照品,经协作标定,候选品中Anti-TpoAb免疫活性均值为434IU/支.结论:本标准品中Anti-TgAb免疫活性定义为4000IU/支,Anti-TpoAb免疫活性定义为430IU/支.本标准品的建立,为国产Anti-TgAb 和Anti-TpoAb试剂盒提供了定量依据,本文还对目前国内常用的几种进口Anti-TgAb 和Anti-TpoAb定量免疫分析试剂盒进行了比较分析,供临床和检验医生参考.     

甲状腺素工作标准品的研制

制备供免疫分析用的甲状腺素工作标准品并标定其免疫效价。用每升含951．22mg T4的40mmol/L NaOH溶液和甲状腺素基础值为零的去激素血清配制浓度为1000μg/L的T4表溶液。将该血清溶液按每安瓿1mL分装（100ng/安瓿）、冻干。以国产T4RIA药盒现行标准品为对照品标定T4工作标准品效价，并从工作标准品的剂量－反应曲线上读出本所制备的1601－9703质控血清的T4测定值。以T4药盒标准品为对照品，工作标准品平均免疫效价为946ng/安瓿，95％概率水平的可信限为915－976ng/安瓿。从工作标准品的剂量－反应曲线上读出低、中、高值质控血清的T4测定值，在以相应的参考值为参照时，测定值的偏倚均小于10％。新制备的T4工作标准品适用于国产T4放射免疫分析药盒标准品的核对工作及其质控血清的标定工作。     

三碘甲腺原氨酸（T3）第三次免疫测定用国家标准品的建立

目的：建立三碘甲腺原氨酸（T3）第三次定量免疫测定用国家标准品。方法和结果：精确称取高纯T3原料,用含0．5％BSA的0．02MPBS缓冲液配制成100ng／ml溶液,经无菌过滤处理后,分装、冻干制成候选品。以T3第二次国家标准品（150550-0004）为对照品,经协作标定,候选品中L免疫活性为50ng／支。结论：候选品各项指标均符合要求,可作为L第三次免疫测定用国家标准品使用。本标准品的建立,为L免疫测定用试剂盒的标准化提供定量基础。     

乙型肝炎病毒表面抗原国家定量标准品的研制

目的 研制乙型肝炎病毒表面抗原(HBsAg)国家定量标准品及定量线性参考品.方法 收集各地乙型肝炎患者和健康献血员血样,用不同厂家乙型肝炎、丙型肝炎病毒血清学试剂盒筛选.6个协作单位用7种试剂以世界卫生组织(WHO)HBsAg定量标准品标定1份HBsAg国家定量标准品,稀释后得到HBsAg国家定量线性参考品.结果经7种试剂共21次协作标定,得到HBsAg国家定量标准品的浓度为1226 IU/ml,各次检测结果的变异系数均小于15%.将国家定量标准品系列稀释后得到由8个不同浓度血清组成的HBsAg国家线性参考品,经检测后表明其浓度与理论浓度的差值均<15%,通过加速试验检测该参考品的稳定性效期暂定为5年.将其作为HBsAg国家线性参考品并初步限定检测结果的允许值范围.结论 标定1份HBsAg国家定量标准品并建立了由8份血清组成的HBsAg国家定量线性参考品.     

癌胚抗原工作标准品的制备

制备供免疫分析用的癌胚抗原工作标准品并标定其免疫效价,用4．6mg/LCEA纯品溶液和PH7．4的1％人白蛋白－50mmol Na2HPO4-NaH2PO4溶液配制浓度为320μg/L的工作标准品溶液。将该溶液按每安瓿（amp.)0.5mL(160ng/amp.)分装、冻干。以国产CEA RIA和CEA IRMA药盒现行标准品为对照品,标定工作标准品效价,并与国际参考制剂1stIRP CEA HUMAN73／601进行对照实验。以药盒标准品为对照品,工作标准品平均免疫效价为163ng/amo.,95%可信为159－168ng/amp.,工作标准品与国际参考剂的剂量－反应曲线不显著偏离平行。CEA工作标准品与国际参考制剂1stIRP CEA73/60在免疫学反应中是同质的,可以作为实验室间对照品使用。     

人垂体促卵泡生成激素免疫测定用标准品研制

随着人垂体促卵泡生成激素（FSH）放射免疫分析及酶联免疫分析技术在临床病理诊断中广泛应用,对其使用的各种测定试剂盒的标准化要求不断提高,必须提供一种准确可靠的标准物质供试剂盒生产者和科研工作使用,为此世界卫生组织（WHO）以高纯的人垂体FSH做原料制备了批号为83/575的标准品,以人垂体FSH、LH第二次国际参考品（批号78/549）为对照品,组织国际协作标定,1986年正式作为国际标准品发放使用,但因此次国际协作标定结果免疫活性结果不均一,所以根据其生物活性标定结果将本标准品定为80IU/支,而83/575的免疫活性标定结果仅为16IU/支,二者相差5倍,造成免疫活性不连续,在国际免疫分析界未得到广泛应用,因而WHO决定FSH IS（83/575）作为生物标准品,78/549仍被用作免疫分析技术标准化工作的参考物质,必须尽快提供一种与IRP（78/549）免疫活性相符的参考物质供试剂盒标准化使用,因而开展了本工作。     

重组人促甲状腺激素在分化型甲状腺癌诊治中的应用

分化型甲状腺癌(DTC)患者常规随访及行131I治疗时,须停用甲状腺素。停用甲状腺素而引发的甲状腺功能减退症状可显著影响DTC患者的生活质量。使用重组人促甲状腺激素(rhTSH)辅助DTC患者的随访及131I治疗,患者不须停用甲状腺素,因此可有效地避免因停用甲状腺素对患者生活质量的影响。本文就rhTSH在DTC诊治中的应用作一综述。     

三碘甲腺原氨酸免疫分析工作标准品的研制

制备供免疫分析用的3,5,3-三碘甲腺原氨酸(T3)工作标准品并标定其免疫效价.用每升含193.27mg T3的40mol/L NaOH溶液和去激素血清,配制浓度为100μg/L的T3溶液.将该溶液按每1mL分装,冻干.以国产现行标准品为对照品,标定配制的T3工作标准品免疫效价为91.4ng/安瓿,95%的可信限为88-95ng/安瓿.并从T3药盒工作标准曲线上,读出本所1601-9703批低、中、高质控血清的T3值.在以相应的参考值为参照时,测定值的偏倚均小于10%.总之,配制的T3工作标准品适用于国产T3RIA药盒标准品的核对及其质控血清的标定.     

The international standard for human interleukin-2. Calibration by international collaborative study

Three lyophilized preparations of interleukin-2 coded 86/500, 86/564 and 86/504 have been evaluated in an international collaborative study for their suitability as an international standard. All of the preparations performed well in the different bioassay systems included in the study, and showed excellent stability on accelerated temperature degradation. Material similar to that in preparation 86/504 has served well as an interim reference reagent for interleukin-2 for 3 years. Therefore with the agreement of the study participants and the authorization of the Expert Committee on Biological Standardization of the World Health Organization, the preparation coded 86/504 was established in 1987 as the 1st international standard for interleukin-2, with a defined potency of 100 IU/ampoule.     

The 2nd International Standard for human granulocyte colony stimulating factor

Five candidate preparations of human sequence recombinant granulocyte-colony stimulating factor (G-CSF) were formulated and lyophilized at NIBSC prior to evaluation in a collaborative study for their suitability to serve as a replacement International Standard (IS). The preparations were tested by 13 laboratories using in vitro bioassays. The candidate preparation 09/136 was judged suitable to serve as a replacement international standard based on the data obtained for biological activity and stability. On the basis of the results reported here, the preparation coded 09/136 was established by the WHO Expert Committee on Biological Standardization (ECBS) as the 2nd IS for human G-CSF with an assigned value for G-CSF activity of 95,000 IU/ampoule. Calibration of the 2nd IS is primarily based on the bioassay in use in various laboratories and relies entirely on the estimates calculated relative to the WHO 1st IS for continuity of the IU.     

Development of monoclonal antibody-based immunoassays for detection of Helicobacter pylori neutrophil-activating protein

One candidate preparation of human sequence recombinant transforming growth factor-β3 (TGF-β3) was formulated and lyophilized at NIBSC prior to evaluation in a collaborative study for its suitability to serve as an international standard. The preparation was tested by 8 laboratories using in vitro bioassays and immunoassays. The candidate preparation 09/234 was judged suitable to serve as an international standard based on the data obtained for biological activity and stability. On the basis of the results reported here, the preparation coded 09/234 was established by the WHO Expert Committee on Biological Standardisation (ECBS) as the WHO 1st IS for human TGF-β3 with an assigned value for TGF-β3 activity of 19,000 IU/ampoule.     

Evaluation of a candidate international standard preparation for human anti-Toxoplasma immunoglobulin G

A freeze-dried human serum preparation containing immunoglobulin G (IgG) to Toxoplasma gondii was assessed for its suitability as an international reference reagent in an international collaborative study by 24 laboratories from 17 countries. This candidate standard was compared with the third international standard (IS) for human anti-Toxoplasma serum, TOXM, with the previous second IS, TOXS, and with a range of other serum samples. Samples were tested with the Sabin-Feldman dye test and a range of agglutination assays and enzyme immunoassays. This study emphasizes the need for appropriate standards if intermethod agreement of estimates is to be achieved. On the basis of the results of this study, the preparation was established by the World Health Organization as the first IS for human anti-Toxoplasma IgG, with an assigned potency of 20 IU per ampoule of total anti-Toxoplasma antibodies.     

Implications for the assay and biological activity of interleukin-8: results of a WHO international collaborative study

Eight ampouled preparations of interleukin-8 (IL-8) have been evaluated for their suitability to serve as an international standard for IL-8 by 30 laboratories in 12 countries in an international collaborative study. The preparations were assayed in a wide range of in vitro bioassays and immunoassays. It is clear from the study that different recombinant preparations of IL-8 can have different biological specific activities, even though all were produced using E. coli. It is of interest that the intra-laboratory variability of estimates provided by several neutrophil degranulation bioassays was less than that of the immunoassays, suggesting that these bioassays can be as precise, if not more so, than immunoassays. In addition, immunoassay estimates of IL-8 preparations differed from those of bioassays, illustrating the fact that immunoassays do not necessarily measure biologically active cytokine. The large reduction in the inter-laboratory variability of estimates in terms of a common reference preparation clearly illustrates the need for a standard for IL-8. On the basis of the results reported here, with the agreement of the participants of the study and with the authorisation of the Expert Committee on Biological Standardization (ECBS) of the World Health Organization (WHO) the preparation of IL-8 (89/520) was established as the International Standard for IL-8 with an assigned unitage of 1000 IU/ampoule.     

The World Health Organization reference reagent for keratinocyte growth factor,KGF

Preparations of human sequence recombinant keratinocyte growth factor (KGF), fibroblast growth factor-7 (FGF-7) synthesized in E. coli were formulated and lyophilized at National Institute for Biological Standards and Control (NIBSC) and evaluated in a collaborative study using in vitro bioassays and immunoassays. One candidate standard was the full-length mature KGF molecule and the two others were different formulations of a truncated form of the molecule, KGF (24-163). The study demonstrated the difficulty of performing interlaboratory comparison of assays without a common reference standard and differences in dose-response relationships between molecular variants. On the basis of the results reported here, the World Health Organization (WHO) established the preparation coded 03/150 as the WHO reference reagent for human KGF, with an assigned unitage of 4000 units of KGF per ampoule and the preparation coded 03/148 as the WHO reference reagent for human KGF (24-163), with an assigned unitage of 9000 units of KGF(24-163) per ampoule. Details on ordering the reference reagents can be found at www.nibsc.ac.uk.     

Gestational Age-specific Cut-off Values Are Needed for Diagnosis of Subclinical Hypothyroidism in Early Pregnancy

During the first trimester of pregnancy, thyroid-stimulating hormone (TSH) >2.5 mIU/L has been suggested as the universal criterion for subclinical hypothyroidism. However, TSH levels change continuously during pregnancy, even in the first trimester. Therefore the use of a fixed cut-off value for TSH may result in a different diagnosis rate of subclinical hypothyroidism according to gestational age. The objective of this study was to obtain the normal reference range of TSH during the first trimester in Korean gravida and to determine the diagnosis rate of subclinical hypothyroidism using the fixed cut-off value (TSH >2.5 mIU/L). The study population consisted of pregnant women who were measured for TSH during the first trimester of pregnancy (n=492) and nonpregnant women (n=984). Median concentration of TSH in pregnant women was lower than in non-pregnant women. There was a continuous decrease of median TSH concentration during the first trimester of pregnancy (median TSH concentration: 1.82 mIU/L for 3+0 to 6+6 weeks; 1.53 mIU/L for 7+0 to 7+6 weeks; and 1.05 mIU/L for 8+0 to 13+6 weeks). Using the fixed cut-off value of TSH >2.5 mIU/L, the diagnosis rate of subclinical hypothyroidism decreased significantly according to the gestational age (GA) at TSH (25% in 3+0 to 6+6 weeks, 13% in 7+0 to 7+6 weeks, and 9% for 8+0 to 13+6 weeks, P<0.001), whereas the diagnosis rate was 5% in all GA with the use of a GA-specific cut-off value (P=0.995). Therefore, GA-specific criteria might be more appropriate for the diagnosis of subclinical hypothyroidism.     

促甲状腺激素生物素-链亲和素TRFIA定量检测法的建立

用Eu3＋标记链亲和素（SA）,分别用两株识别人促甲状腺素（human thyroid stimulating hormone,hTSH）的单克隆抗体包被96微孔板并生物素化,建立了高灵敏的hTSH生物素-链亲和素（BAS）时间分辨荧光免疫分析（TRFIA）检测法。本法的灵敏度为0.02mIU/L;批内CV为2.7%~3.5%,批间CV为3.2%~3.8%;平均回收率为100.26%;与电化学发光免疫分析技术（ECLIA）比对,相关系数达0.9544,线性方程为Y=1.3133X-32.297;与ECLIA临床测定值也呈明显相关;正常值范围为0.33~3.09mIU/L。本文建立的hTSH-BAS-TRFIA具有灵敏度高,特异性强,稳定性好,测定范围宽,检测时间短和非放射性等优点,具有良好的应用价值。     

The World Health Organization reference reagent for keratinocyte growth factor, KGF

Preparations of human sequence recombinant keratinocyte growth factor (KGF), fibroblast growth factor-7 (FGF-7) synthesized in E. coli were formulated and lyophilized at National Institute for Biological Standards and Control (NIBSC) and evaluated in a collaborative study using in vitro bioassays and immunoassays. One candidate standard was the full-length mature KGF molecule and the two others were different formulations of a truncated form of the molecule, KGF (24-163). The study demonstrated the difficulty of performing interlaboratory comparison of assays without a common reference standard and differences in dose-response relationships between molecular variants. On the basis of the results reported here, the World Health Organization (WHO) established the preparation coded 03/150 as the WHO reference reagent for human KGF, with an assigned unitage of 4000 units of KGF per ampoule and the preparation coded 03/148 as the WHO reference reagent for human KGF (24-163), with an assigned unitage of 9000 units of KGF(24-163) per ampoule.