title: Serogroup-specific bacterial engineered glycoproteins as novel antigenic targets for diagnosis of shiga toxin-producing-escherichia coli-associated hemolytic-uremic syndrome.
abstract: Human infection with Shiga toxin-producing Escherichia coli (STEC) is a major cause of postdiarrheal hemolytic-uremic syndrome (HUS), a life-threatening condition characterized by hemolytic anemia, thrombocytopenia, and acute renal failure. E. coli O157:H7 is the dominant STEC serotype associated with HUS worldwide, although non-O157 STEC serogroups can cause a similar disease. The detection of anti-O157 E. coli lipopolysaccharide (LPS) antibodies in combination with stool culture and detection of free fecal Shiga toxin considerably improves the diagnosis of STEC infections. In the present study, we exploited a bacterial glycoengineering technology to develop recombinant glycoproteins consisting of the O157, O145, or O121 polysaccharide attached to a carrier protein as serogroup-specific antigens for the serological diagnosis of STEC-associated HUS. Our results demonstrate that using these antigens in indirect ELISAs (glyco-iELISAs), it is possible to clearly discriminate between STEC O157-, O145-, and O121-infected patients and healthy children, as well as to confirm the diagnosis in HUS patients for whom the classical diagnostic procedures failed. Interestingly, a specific IgM response was detected in almost all the analyzed samples, indicating that it is possible to detect the infection in the early stages of the disease. Additionally, in all the culture-positive HUS patients, the serotype identified by glyco-iELISAs was in accordance with the serotype of the isolated strain, indicating that these antigens are valuable not only for diagnosing HUS caused by the O157, O145, and O121 serogroups but also for serotyping and guiding the subsequent steps to confirm diagnosis.
authors.affiliation: Instituto de Investigaciones Biotecnológicas Dr. Rodolfo A. Ugalde, Instituto Tecnológico de Chascomús (IIB-INTECH), Universidad Nacional de San Martín, CONICET, San Martín, Buenos Aires, Argentina.
authors.forename: Luciano J
authors.initials: LJ
authors.lastname: Melli
authors.affiliation: Instituto de Investigaciones Biotecnológicas Dr. Rodolfo A. Ugalde, Instituto Tecnológico de Chascomús (IIB-INTECH), Universidad Nacional de San Martín, CONICET, San Martín, Buenos Aires, Argentina.
authors.forename: Andrés E
authors.initials: AE
authors.lastname: Ciocchini
authors.affiliation: Instituto de Investigaciones Biotecnológicas Dr. Rodolfo A. Ugalde, Instituto Tecnológico de Chascomús (IIB-INTECH), Universidad Nacional de San Martín, CONICET, San Martín, Buenos Aires, Argentina.
authors.forename: Ana J
authors.initials: AJ
authors.lastname: Caillava
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Canada.
authors.forename: Nicolás
authors.initials: N
authors.lastname: Vozza
authors.affiliation: Servicio Fisiopatogenia, Instituto Nacional de Enfermedades Infecciosas-ANLIS Dr. Carlos G. Malbrán, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina.
authors.forename: Isabel
authors.initials: I
authors.lastname: Chinen
authors.affiliation: Servicio Fisiopatogenia, Instituto Nacional de Enfermedades Infecciosas-ANLIS Dr. Carlos G. Malbrán, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina.
authors.forename: Marta
authors.initials: M
authors.lastname: Rivas
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Canada.
authors.forename: Mario F
authors.initials: MF
authors.lastname: Feldman
authors.affiliation: Instituto de Investigaciones Biotecnológicas Dr. Rodolfo A. Ugalde, Instituto Tecnológico de Chascomús (IIB-INTECH), Universidad Nacional de San Martín, CONICET, San Martín, Buenos Aires, Argentina jugalde@iibintech.com.ar dcomerci@iibintech.com.ar.
authors.forename: Juan E
authors.initials: JE
authors.lastname: Ugalde
authors.affiliation: Instituto de Investigaciones Biotecnológicas Dr. Rodolfo A. Ugalde, Instituto Tecnológico de Chascomús (IIB-INTECH), Universidad Nacional de San Martín, CONICET, San Martín, Buenos Aires, Argentina Comisión Nacional de Energía Atómica, Grupo Pecuario, Centro Atómico Ezeiza, Buenos Aires, Argentina jugalde@iibintech.com.ar dcomerci@iibintech.com.ar.
authors.forename: Diego J
authors.initials: DJ
authors.lastname: Comerci
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology*; D006463:Hemolytic-Uremic Syndrome / Q000175:diagnosis*; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood; D007223:Infant; D007231:Infant, Newborn; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012189:Retrospective Studies; D012703:Serotyping / Q000379:methods*; D054323:Shiga-Toxigenic Escherichia coli / Q000276:immunology*; D016037:Single-Blind Method
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D006023:Glycoproteins; D007075:Immunoglobulin M; D011994:Recombinant Proteins