Fluorescent multivalent opsonophagocytic assay for measurement of functional antibodies to Streptococcus pneumoniae.
We developed fluorescent mono- and multivalent opsonophagocytic assays (fOPA and fmOPA, respectively) specific for seven Streptococcus pneumoniae serotypes (4, 6B, 9V, 14, 18C, 19F, and 23F). Bacterial survival was quantitated with alamar blue, a fluorescent metabolic indicator. Both fOPA and fmOPA allow for determination of viability endpoints for up to seven serotypes with high levels of agreement to the reference method. The fmOPA eliminates colony counting, reduces serum volume, and produces results in 1 day.
Bieging
Kathryn T
KT
Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA 30333, USA.
Rajam
Gowrisankar
G
Holder
Patricia
P
Udoff
Ross
R
Carlone
George M
GM
Romero-Steiner
Sandra
S
D000907:Antibodies, Bacterial / Q000097:blood*; D015169:Colony Count, Microbial; D006801:Humans; D009895:Opsonin Proteins; D010078:Oxazines; D010587:Phagocytosis; D011008:Pneumococcal Infections / Q000175:diagnosis*; D012703:Serotyping / Q000379:methods; D013296:Streptococcus pneumoniae / Q000145:classification / Q000166:cytology / Q000276:immunology*; D014966:Xanthenes
D000907:Antibodies, Bacterial; D009895:Opsonin Proteins; D010078:Oxazines; D014966:Xanthenes; C005843:resazurin