Exploring of pH conditions for HIV detection(ELISA) in IVIG finished products
10.13200/j.cnki.cjb.004379
- VernacularTitle: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
- Author:
ZHENG Xiaobei
- Publication Type:Journal Article
- Keywords:
Human immunoglobulin(pH 4) for intravenous injection(IVIG);
human immunodeficiency virus(HIV);
pH;
Cutoff value;
Tris-HCl solution
- From:
Chinese Journal of Biologicals
2024;37(12):1463-1469
- CountryChina
- Language:Chinese
-
Abstract:
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. 5-8. 0 is more suitable for HIV detection in IVIG. 1 mol/L Tris-HCl solution was used to adjust the pH of IVIG to 7. 5-8. 0 for HIV detection(ELISA)in IVIG, which effectively solves the problem that the pH bias in IVIG finished product is not conducive to HIV detection, with no effect on the detection volume of the finished product, thus significantly improving the accuracy and reliability of the detection results.
