title: Comparative study of different sources of pertussis toxin (PT) as coating antigens in IgG anti-PT enzyme-linked immunosorbent assays.
abstract: In an effort to improve the reliability and reproducibility of serological assays for Bordetella pertussis, a collaborative study was conducted to compare four different sources of pertussis toxin (PT) as coating antigens in the immunoglobulin G (IgG) anti-PT enzyme-linked immunosorbent assay (ELISA). Four sources of PT were used as coating antigens in the IgG anti-PT ELISA in four different testing laboratories (labs A to D) to determine whether the different antigen preparations and different laboratories influenced assay results. A panel of 60 sera consisting of deidentified human specimens from previous vaccination trials of healthy adults and infants and clinical specimens from outbreak settings was tested. In the four laboratories, each sample was tested three times with the four PT antigens according to the standard coating optimization and IgG anti-PT ELISA testing procedures used in that laboratory. Differences among the antigens, as well as intra- and interlaboratory variability, were evaluated. Excellent agreement was observed with the test panel results among the four antigens within each laboratory. Concordance correlation coefficient (r(c)) measurements among the different antigens ranged from 0.99, 0.99 to 1.00, 1.00, and 0.97 to 1.00 for labs A to D, respectively. The comparisons between pairs of laboratories also indicated a high degree of concordance for each PT preparation, with r(c) measurements between 0.90 and 0.98, 0.93 and 0.99, 0.92 and 0.98, and 0.93 and 0.99 for antigens 1 to 4, respectively. Relatively minor differences in results were observed among laboratories or among antigens, suggesting that the four PT antigens are quite similar and could be considered for acceptance in harmonized immunoassays used for serodiagnosis or vaccine evaluation.
authors: Division of Bacterial Diseases, NCIRD, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors: Aditi
authors: A
authors: Kapasi
authors: Bruce D
authors: BD
authors: Meade
authors: Brian
authors: B
authors: Plikaytis
authors: Lucia
authors: L
authors: Pawloski
authors: Monte D
authors: MD
authors: Martin
authors: Sandra
authors: S
authors: Yoder
authors: Michael T
authors: MT
authors: Rock
authors: Séverine
authors: S
authors: Coddens
authors: Valérie
authors: V
authors: Haezebroeck
authors: Françoise
authors: F
authors: Fievet-Groyne
authors: Garvin
authors: G
authors: Bixler
authors: Charles
authors: C
authors: Jones
authors: Stephen
authors: S
authors: Hildreth
authors: Kathryn M
authors: KM
authors: Edwards
authors: Nancy E
authors: NE
authors: Messonnier
authors: Maria L
authors: ML
authors: Tondella
mesh_terms: D000328:Adult; D000907:Antibodies, Bacterial / Q000097:blood*; D001886:Bordetella pertussis / Q000276:immunology*; D002675:Child, Preschool; D019411:Clinical Laboratory Techniques / Q000379:methods* / Q000592:standards; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D007223:Infant; D037342:Pertussis Toxin; D015203:Reproducibility of Results; D014917:Whooping Cough / Q000175:diagnosis* / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D007074:Immunoglobulin G; D037342:Pertussis Toxin