Identification of Pneumococcal Serotypes by PCR-Restriction Fragment Length Polymorphism
| Autor: | Jose Yuste, Irene González-Rodríguez, Fernando Vázquez, María del Mar García-Suárez, Enrique Santiago, María Dolores Cima-Cabal |
|---|---|
| Přispěvatelé: | University of Oviedo (España), Ministerio de Educación, Cultura y Deporte (España), Universidad Internacional de La Rioja (España), Universidad de Oviedo, Ministerio de Educación y Cultura (España), Universidad Internacional de La Rioja |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Serotype 030106 microbiology Clinical Biochemistry Biology medicine.disease_cause Article DNA sequencing Restriction fragment 03 medical and health sciences PCR-RFLP 0302 clinical medicine Streptococcus pneumoniae medicine 030212 general & internal medicine Typing serotype pcr-rflp streptococcus pneumoniae Genetics lcsh:R5-920 Restriction enzyme biology.protein Quellung reaction Restriction fragment length polymorphism lcsh:Medicine (General) |
| Zdroj: | Repisalud Instituto de Salud Carlos III (ISCIII) Digital.CSIC. Repositorio Institucional del CSIC instname Scopus RUO. Repositorio Institucional de la Universidad de Oviedo Diagnostics, Vol 9, Iss 4, p 196 (2019) Diagnostics Volume 9 Issue 4 RUO: Repositorio Institucional de la Universidad de Oviedo Universidad de Oviedo (UNIOVI) |
| Popis: | 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&ndash wzh&ndash wzd&ndash 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|