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Clinical and Diagnostic Laboratory Immunology, October 2005, p. 1238-1242, Vol. 12, No. 10
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.10.1238-1242.2005

Fluorescent Multivalent Opsonophagocytic Assay for Measurement of Functional Antibodies to Streptococcus pneumoniae

Kathryn T. Bieging, Gowrisankar Rajam, Patricia Holder, Ross Udoff, George M. Carlone, and Sandra Romero-Steiner*

Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333

Received 16 June 2005/ Returned for modification 11 July 2005/ Accepted 15 August 2005

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.

* Corresponding author. Mailing address: MS A-36, Respiratory Diseases Immunology Section, Respiratory Diseases Branch, Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA 30333. Phone: (404) 639-2473. Fax: (404) 639-3115. E-mail: Ssteiner{at}cdc.gov.

Clinical and Diagnostic Laboratory Immunology, October 2005, p. 1238-1242, Vol. 12, No. 10
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.10.1238-1242.2005






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