Identification of Pneumococcal Serotypes by PCR-Restriction Fragment Length Polymorphism

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...

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Bibliographic Details
Published in:Diagnostics (Basel) 2019-11, Vol.9 (4), p.196
Main Authors: García-Suárez, María Del Mar, González-Rodríguez, Irene, Cima-Cabal, María Dolores, Yuste, Jose Enrique, Vazquez, Fernando, Santiago, Enrique
Format: Article
Language:English
Subjects:
Enzymes
pcr-rflp
Population
serotype
Streptococcus infections
streptococcus pneumoniae
Vaccines
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Summary: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 B and A genes were compiled, and a computer screening of restriction enzymes was performed. The region and 9I 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.
ISSN:2075-4418
2075-4418
DOI:10.3390/diagnostics9040196