Effects of hemolysis, chylemia and repeated freeze-thawing of human serum immunized with pneumococcal vaccine on ELISA clinical evaluation method
10.13200/j.cnki.cjb.004500
- VernacularTitle:肺炎链球菌疫苗免疫后人血清中溶血、乳糜血以及反复冻融对ELISA临床评价方法的影响
- Author:
SHI Gang
- Publication Type:Journal Article
- Keywords:
Pneumococcal vaccine;
ELISA;
Freeze-thawing;
Hemolysis;
Chylemia;
IgG antibody
- From:
Chinese Journal of Biologicals
2025;38(06):705-712
- CountryChina
- Language:Chinese
-
Abstract:
Objective To confirm the effects of hemolysis,chylemia and repeated freeze-thawing of human serum immunized with pneumococcal vaccine on ELISA for the detection of specific IgG antibody against pneumococcal capsular polysaccharide,and to promote the continuous development and improvement of the evaluation methods for clinical serum IgG antibody content of pneumococcal vaccine.Methods The same batch of reconstituted laboratory internal quality control serum 09CS was aliquoted into different volumes and subjected to 1 to 12 freeze-thaw cycles.The IgG antibodies of 24 types were detected respectively with the serum after single freeze-thaw and the repeated freeze-thaw serum.The ELISA test for detecting the content of IgG antibodies specific to pneumococcal capsular polysaccharides was carried out,and the test results were systematically analyzed.The laboratory internal quality control serum 09CS was respectively mixed with hemolyzed and chylous serum after immunization in different volume fractions to prepare hemolyzed and chylous serum,and the IgG antibodies of 24 types were detected respectively.The test was repeated three times,and the recovery rate was calculated.Results The repeated freeze-thaw serum and single freeze-thaw serum were detected for 24 types respectively,and the detection results showed good linear relationship with R~2> 0.99.The detection results were analyzed by Lin's concordance correlation coefficient(r_c),and the r_c was 0.992 3(95% CI:0.990 2-0.993 9),indicating that the test results of single freeze-thaw serum and repeated freeze-thaw serum were highly consistent.The accuracy of serum recovery rate was74.65% for hemolyzed serum and 70.48% for chylous serum,both in line with the acceptance range of accuracy±40%.Conclusion During the ELISA test for the specific IgG antibody content against pneumococcal capsular polysaccharide,hemolyzed and chylous serum and repeated freeze-thawing of serum for 12 times have no effect on the detection,which can satisfy the detection requirements for clinical serum antibody evaluation of pneumococcal vaccine.
