title: A Novel IgM-capture enzyme-linked immunosorbent assay using recombinant Vag8 fusion protein for the accurate and early diagnosis of Bordetella pertussis infection.
abstract: An ELISA that measures anti-PT IgG antibody has been used widely for the serodiagnosis of pertussis; however, the IgG-based ELISA is inadequate for patients during the acute phase of the disease because of the slow response of anti-PT IgG antibodies. To solve this problem, we developed a novel IgM-capture ELISA that measures serum anti-Bordetella pertussis Vag8 IgM levels for the accurate and early diagnosis of pertussis. First, we confirmed that Vag8 was highly expressed in all B. pertussis isolates tested (n = 30), but little or none in other Bordetella species, and that DTaP vaccines did not induce anti-Vag8 IgG antibodies in mice (i.e. the antibody level could be unaffected by the vaccination). To determine the immune response to Vag8 in B. pertussis infection, anti-Vag8 IgM levels were compared between 38 patients (acute phase of pertussis) and 29 healthy individuals using the anti-Vag8 IgM-capture ELISA. The results revealed that the anti-Vag8 IgM levels were significantly higher in the patients compared with the healthy individuals (P < 0.001). ROC analysis also showed that the anti-Vag8 IgM-capture ELISA has higher diagnostic accuracy (AUC, 0.92) than a commercial anti-PT IgG ELISA kit. Moreover, it was shown that anti-Vag8 IgM antibodies were induced earlier than anti-PT IgG antibodies on sequential patients' sera. These data indicate that our novel anti-Vag8 IgM-capture ELISA is a potentially useful tool for making the accurate and early diagnosis of B. pertussis infection.
authors.affiliation: Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo.
authors.forename: Nao
authors.initials: N
authors.lastname: Otsuka
authors.affiliation: Department of Pediatrics, Konan Kosei Hospital, Konan.
authors.forename: Kensei
authors.initials: K
authors.lastname: Gotoh
authors.affiliation: Department of Pediatrics, Konan Kosei Hospital, Konan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Nishimura
authors.affiliation: Department of Pediatrics, Konan Kosei Hospital, Konan.
authors.forename: Takao
authors.initials: T
authors.lastname: Ozaki
authors.affiliation: Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo.|Department of Pediatrics, St. Marianna University School of Medicine, Kawasaki.
authors.forename: Yukitsugu
authors.initials: Y
authors.lastname: Nakamura
authors.affiliation: Denka Seiken Co. Ltd, Tokyo.
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Haga
authors.affiliation: Denka Seiken Co. Ltd, Tokyo.
authors.forename: Makoto
authors.initials: M
authors.lastname: Yamazaki
authors.affiliation: Denka Seiken Co. Ltd, Tokyo.
authors.forename: Fumio
authors.initials: F
authors.lastname: Gondaira
authors.affiliation: Section of Pediatrics, Department of Medicine, Division of Oral & Medical Management, Fukuoka Dental College, Fukuoka.
authors.forename: Kenji
authors.initials: K
authors.lastname: Okada
authors.affiliation: Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo.|Department of Pediatrics, St. Marianna University School of Medicine, Kawasaki.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Miyaji
authors.affiliation: Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Toyoizumi-Ajisaka
authors.affiliation: Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo.
authors.forename: Keigo
authors.initials: K
authors.lastname: Shibayama
authors.affiliation: Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo.|Department of Bacteriology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Yoshichika
authors.initials: Y
authors.lastname: Arakawa
authors.affiliation: Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo.
authors.forename: Kazunari
authors.initials: K
authors.lastname: Kamachi
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001886:Bordetella pertussis / Q000276:immunology*; D042241:Early Diagnosis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood*; D011506:Proteins; D012372:ROC Curve; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012698:Serologic Tests / Q000379:methods*; D014917:Whooping Cough / Q000175:diagnosis*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007075:Immunoglobulin M; D011506:Proteins; D011994:Recombinant Proteins
keywords: Bordetella pertussis; IgM-capture enzyme-linked immunosorbent assay (ELISA); Vag8; serodiagnosis