Standardization of a chemiluminescence method for the measurement of meningococcal opsonins using ethanol fixed...

A chemiluminescence (CL) method using polymorphonuclear leukocytes (PMNLs) and an automatic photoluminometer was used to measure serum opsonins to viable and inactivated group B meningococci. Continuous mixing at 37 degrees C both during opsonization and phagocytosis was essential for optimal CL responses. The CL response increased rapidly during an opsonization time up to 7.5 min, and with PMNL and bacteria concentrations up to 37.5 X 10(5) and 3.8 X 10(7) cells/ml, respectively. Opsonized ethanol fixed meningococci gave CL responses similar to those of viable meningococci, but had a better reproducibility. Using the ethanol fixed bacteria, the variation of PMNLs from different donors, the day-to-day variation, and the coefficient of variation of the CL responses, were all less than 10%. The opsonic activity of convalescent sera from 10 patients with meningococcal disease was markedly higher than that of sera obtained during the acute phase of the disease. Thus, this standardized CL assay using ethanol fixed bacteria is a highly reproducible and sensitive method for measuring serum opsonins to meningococci.