Identification of Pneumococcal Serotypes by PCR–Restriction Fragment Length Polymorphism
Autor:
García-Suárez, María del Mar (1)
González-Rodríguez, Irene
Cima-Cabal, María Dolores (1)
Yuste, Jose Enrique
Vazquez, Fernando
Santiago, Enrique
Fecha:
18/11/2019Palabra clave:
Tipo de Ítem:
Articulo Revista IndexadaDirección web:
https://www.mdpi.com/2075-4418/9/4/196
Resumen:
Streptococcus pneumoniae shows more than 90 capsular serotypes that can be distinguished by their reactivity against antisera. The main objective of this work was the development of a molecular method for serotyping without the use of antisera. A computer program containing an algorithm was used to search in a database for potentially useful enzymes for Restriction Fragment Length Polymorphism-RFLP typing, in order to maximize the discrimination between different serotypes. DNA sequences of 90 serotypes for the region between dexB and aliA genes were compiled, and a computer screening of restriction enzymes was performed. The wzg–wzh–wzd–wze region and Sse9I restriction predicted unique PCR-RFLP patterns for 39 serotypes and eight serogroups. A second restriction enzyme resolved fragment specific patterns for 25 serotypes. The method was tested with 98 serotype-unknown clinical isolates. PCR-RFLP analysis deduced correct serotypes that were confirmed by Quellung reaction for 78.5% of the isolates.
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