静注人免疫球蛋白(pH 4)成品中人类免疫缺陷病毒检测(ELISA法)pH条件的摸索  

Exploring of pH conditions for HIV detection(ELISA)in IVIG finished products

作　　者：郑宵蓓 王月 邓琨 龚钦 成立 周志军 郭佳茹 杨轶 童剑啸 柯兵兵 张金 ZHENG Xiaobei;WANG Yue;DENG Kun;GONG Qin;CHENG Li;ZHOU Zhijun;GUO Jiaru;YANG Yi;TONG Jianxiao;KE Bingbing;ZHANG Jin(Sinopharm Wuhan Biopharmaceuticals Co.,Ltd.,Wuhan 430207,Hubei Province,China;不详)

机构地区：[1]国药集团武汉生物制药有限公司,湖北武汉430207 [2]湖北省药品监督管理局检查中心(湖北省疫苗检查中心),湖北武汉430064 [3]湖北省药品监督检验研究院,湖北武汉430075

出　　处：《中国生物制品学杂志》2024年第12期1463-1469,共7页Chinese Journal of Biologicals

基　　金：湖北省科技厅自然科学基金科研课题(2022CFB521)。

摘　　要：目的 对静注人免疫球蛋白(pH 4)[human immunoglobulin(pH 4)for intravenous injection,IVIG,简称静丙]成品中人类免疫缺陷病毒(human immunodeficiency virus,HIV)检测(ELISA法)的pH条件进行摸索,以期控制静丙HIV的残余风险,保证血液制品的安全性。方法 使用不同pH缓冲液稀释HIV抗体标准物质及抗原标准物质至检测限水平,观察试剂盒对不同pH、检测限水平HIV标准物质的检出情况,进而得出试剂盒适宜pH检出范围。使用不同浓度的Tris-HCl溶液调节静丙成品至试剂盒适宜pH范围,然后稀释HIV抗体标准物质及抗原标准物质至检测限水平,获得静丙适宜的pH检出范围。结果 HIV抗体标准物质和HIV抗原标准物质检测缓冲液pH 7.0~9.0组的A450/630值与生理盐水(NS)组相近,所用HIV试剂盒适宜pH范围为7.0~9.0。0.1 mol/L Tris-HCl溶液稀释静丙4.0至试剂盒适宜pH范围作为稀释液,抗体0.25 NCU/mL水平在静丙pH 7.0~8.0组检测A450/630值高于cutoff值,且明显高于静丙pH 4.0组,0.1 mol/L Tris-HCl溶液加入量占静丙体积1/5以上。1 mol/L Tris-HCl溶液稀释静丙4.0至试剂盒适宜pH范围作为稀释液,抗体0.25 NCU/mL和抗原1.5 U/mL水平在静丙pH 7.0~9.0组检测A450/630值高于cutoff值,1 mol/L Tris-HCl溶液加入量仅为检测总体积的1/28~1/37,对待检样品的有效检测含量几乎不产生影响。结论 相较于pH 4.0的检测条件,pH 7.5~8.0更适用于静丙的HIV检测。确定采用1 mol/L Tris-HCl溶液调节静丙pH至7.5~8.0用于静丙HIV检测(ELISA法)。有效解决了静丙成品pH偏酸不利于HIV检出的问题,同时未对静丙成品的检测体积产生影响,显著提升了检测结果的准确性和可靠性。Objective To explore the pH conditions for human immunodeficiency virus(HIV)detection(ELISA)in the finished products of human immunoglobulin(pH 4)for intravenous injection(IVIG),in order to control the residual HIV risk of IVIG and ensure the safety of blood products.Methods The HIV antibody reference materials and antigen reference materials were diluted to the detection limit level using different pH buffer solutions.The detection of HIV reference materials at different pH and detection limit levels by the reagent kit were observed to determine the appropriate pH detection range of the reagent kit.The pH range of the finished product was adjusted to the appropriate range of the reagent kit by using different concentrations of Tris-HCl solutions,and then the HIV antibody reference materials and HIV antigen reference materials were diluted to the detection limit level to obtain the appropriate pH detection range of the product.Results The A450/630 values of the HIV antibody reference materials and HIV antigen reference materials with detection buffer at pH 7.0-9.0 were similar to those of the normal saline(NS)group,and the suitable pH range for the HIV kit used was pH 7.0-9.0.IVIG 4.0 was diluted with0.1 mol/L Tris-HCl solution to the appropriate pH range of the reagent kit as the diluent.The A450/630 value of antibody 0.25 NCU/mL in IVIG pH 7.0-8.0 group was higher than the cutoff value and significantly higher than that in the IVIG pH 4.0 group.The amount of 0.1 mol/L Tris-HCl solution added accounted for more than 1/5 of the IVIG volume.IVIG 4.0 was diluted with 1 mol/L Tris-HCl solution to the appropriate pH range of the reagent kit as the diluent.The A450/630value of antibody 0.25 NCU/mL and antigen 1.5 U/mL levels was higher than the cutoff value in the IVIG pH 7.0-9.0 group.The amount of 1 mol/L Tris-HCl solution added was only 1/28-1/37 of the total detection volume,and had little effect on the effective detection content of the test sample.Conclusion Compared to the detection conditions of pH 4.0,pH 7.5-

关 键 词：静注人免疫球蛋白(pH 4) 人类免疫缺陷病毒 PH CUTOFF值 Tris-HCl溶液 

分 类 号：R979.5[医药卫生—药品]