A sensitive chemiluminescence based immunoassay for antibody to staphylococcal peptidoglycan.
A sensitive chemiluminescence based immunoassay is described for measuring antibody to staphylococcal peptidoglycan in blood and dialysates from patients undergoing continuous ambulatory peritoneal dialysis (CAPD). Peptidoglycan was isolated from a strain of S. epidermidis obtained from the dialysate of a CAPD patient with peritonitis and after sonication used to coat polystyrene beads. The coated beads were incubated with standard or sample and bound IgG was detected by the addition of affinity-purified goat anti-human IgG labelled with acridinium ester. After a wash stage 0.1 M nitric acid containing 0.1% hydrogen peroxide was added to the beads. Subsequently the chemiluminescence produced following the addition of 0.3 M sodium hydroxide was measured over a 2 s time interval with an automatic luminescence analyser. Using this technique the optimum dilution of serum for detecting antibodies to peptidoglycan was found to be 1/800 and for overnight effluent from CAPD patients the dilution was 1/8. Initial values of serum and dialysate antibody levels from 34 subjects are presented. This method has the advantage that it will detect concentrations of anti-peptidoglycan which are less than 1% of those in sera, the reagents remain stable for long periods and large numbers of samples can be processed on the same day.
Minors
S
S
KRUF Institute of Renal Disease, Cardiff Royal Infirmary, U.K.
Horton
J K
JK
Weeks
I
I
Davies
M
M
Coles
G A
GA
D000907:Antibodies, Bacterial / Q000032:analysis*; D015314:Dialysis Solutions / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000032:analysis; D008163:Luminescent Measurements; D010457:Peptidoglycan / Q000276:immunology* / Q000302:isolation & purification; D010531:Peritoneal Dialysis, Continuous Ambulatory; D013212:Staphylococcus epidermidis / Q000276:immunology*
D000907:Antibodies, Bacterial; D015314:Dialysis Solutions; D007074:Immunoglobulin G; D010457:Peptidoglycan