PCR-Based Method for Shigella flexneri Serotyping: International Multicenter Validation

• Autores: Brengi, Silvina P.; Sun, Qiangzheng; Bolaños, Hilda; Duarte, Francisco; Jenkins, Claire; Pichel, Mariana; Shahnaij, Mohammad; Sowers, Evangeline G; Strockbine, Nancy; Talukder, Kaisar A; Derado, Gordana; Viñas, María R.; Kam, Kai Man; Xu, Jianguo
• Año de publicación: 2019
• Idioma: inglés
• Tipo de recurso: artículo
• Fuente: Journal of Clinical Virology 2019;57(4):e01592-18
• Materia: Reacción en Cadena de la Polimerasa; Shigella flexneri; Serogrupo
• Institución: Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"

Fil: Brengi, Silvina P. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Enterobacterias; Argentina.
Fil: Sun, Qiangzheng. Chinese Center for Disease Control and Prevention. National Institute for Communicable Diseases Control and Prevention. State Key Laboratory for Infectious Disease Prevention and Control, Changping, Beijing; China.
Fil: Bolaños, Hilda. Centro Nacional de Referencia de Bacteriología, Inciensa, Cartago; Costa Rica.
Fil: Duarte, Francisco. Centro Nacional de Referencia de Bacteriología, Inciensa, Cartago; Costa Rica.
Fil: Jenkins, Claire. Public Health England. Gastrointestinal Bacteria Reference Unit, Londres, Inglaterra.
Fil: Pichel, Mariana. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Enterobacterias; Argentina.
Fil: Shahnaij, Mohammad. International Center for Diarrheal Disease Research. Laboratory Sciences and Services Division. Enteric and Food Microbiology Laboratory Dhaka; Bangladés.
Fil: Sowers, Evangeline G. Centers for Disease Control and Prevention. National Institute for Escherichia and Shigella Reference Unit, Atlanta, Georgia, Estados Unidos.
Fil: Strockbine, Nancy. Centers for Disease Control and Prevention. National Institute for Escherichia and Shigella Reference Unit, Atlanta, Georgia, Estados Unidos.
Fil: Talukder, Kaisar A. International Center for Diarrheal Disease Research. Laboratory Sciences and Services Division. Enteric and Food Microbiology Laboratory Dhaka; Bangladés.
Fil: Derado, Gordana. Centers for Disease Control and Prevention. Biostatistics and Information Management Office. Waterborne and Environmental Diseases. Division of Foodborne, Atlanta, Georgia, Estados Unidos.
Fil: Viñas, María R. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Enterobacterias; Argentina.
Fil: Kam, Kai Man. The Chinese University of Hong Kong. Faculty of Medicine. School of Public Health and Primary Care. Stanley Ho Centre for Emerging Infectious Diseases, Hong Kong; China.
Fil: Xu, Jianguo. Chinese Center for Disease Control and Prevention. National Institute for Communicable Diseases Control and Prevention. State Key Laboratory for Infectious Disease Prevention and Control, Changping, Beijing; China.
Shigella spp. are a leading cause of human diarrheal disease worldwide, with Shigella flexneri being the most frequently isolated species in developing countries. This serogroup is presently classified into 19 serotypes worldwide. We report here a multicenter validation of a multiplex-PCR-based strategy previously developed by Q. Sun, R. Lan, Y. Wang, A. Zhao, et al. (J Clin Microbiol 49:3766-3770, 2011) for molecular serotyping of S. flexneri This study was performed by seven international laboratories, with a panel of 71 strains (researchers were blind to their identity) as well as 279 strains collected from each laboratory's own local culture collections. This collaborative work found a high extent of agreement among laboratories, calculated through interrater reliability (IRR) measures for the PCR test that proved its robustness. Agreement with the traditional method (serology) was also observed in all laboratories for 14 serotypes studied, while specific genetic events could be responsible for the discrepancies among methodologies in the other 5 serotypes, as determined by PCR product sequencing in most of the cases. This work provided an empirical framework that allowed the use of this molecular method to serotype S. flexneri and showed several advantages over the traditional method of serological typing. These advantages included overcoming the problem of availability of suitable antisera in testing laboratories as well as facilitating the analysis of multiple samples at the same time. The method is also less time-consuming for completion and easier to implement in routine laboratories. We recommend that this PCR be adopted, as it is a reliable diagnostic and characterization methodology that can be used globally for laboratory-based shigella surveillance.