Validation of colorimetric assay to detect complement-mediated antibody-dependent bactericidal activity against serogroups B and C Neisseria meningitidis.
Colorimetric serum bactericidal assay (cSBA), based on the addition of glucose and a pH indicator to the culture medium after the bactericidal reaction, was validated. The precision measured as repeatability, intermediated precision, and reproducibility was determined as a percentage in titer coincidence between replicas >/=50. Moreover the use of the freeze-dried complement was evaluated in comparison to the traditionally stored by freezing. The results were the following: precision: titer +/-1 two-fold dilution (except for the highly positive serum against serogroup B, where there was titer +/-2 dilutions) percentage in titer coincidence: >/=50. The known titer or +/-1 two-fold dilution was found in the sera titrated with the complement either frozen or freeze-dried. Concluding, cSBA showed to be highly precise, allowing also the use of freeze-dried complement which is another important advantage for this kind of assay.
Rodríguez
Tamara
T
Department of Immunology, Direction of Bacterial Vaccines and Direction of Control of Quality, Finlay Institute, 27 Ave, b/198 and 202, PO Box 16017, La Lisa, Havana, Cuba. trodriguez@finlay.edu.cu
Lastre
Miriam
M
Cedré
Bárbara
B
Fajardo
Esther María
EM
del Campo
Judith
J
Delgado
Ileana
I
Sierra
Gustavo
G
Pérez
Oliver
O
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D001770:Blood Bactericidal Activity; D015941:Complement Hemolytic Activity Assay; D003165:Complement System Proteins / Q000276:immunology*; D006801:Humans; D008589:Meningococcal Infections / Q000276:immunology*; D038541:Neisseria meningitidis, Serogroup B / Q000276:immunology*; D038542:Neisseria meningitidis, Serogroup C / Q000276:immunology*; D011817:Rabbits; D015203:Reproducibility of Results
D000907:Antibodies, Bacterial; D003165:Complement System Proteins