Use of the Agilent 2100 bioanalyzer for rapid and reproducible molecular typing of Streptococcus pneumoniae

Hathaway, Lucy J; Brugger, Silvio D.; Martynova, Alina; Aebi, Suzanne; Mühlemann, Kathrin (2007). Use of the Agilent 2100 bioanalyzer for rapid and reproducible molecular typing of Streptococcus pneumoniae. Journal of clinical microbiology, 45(3), pp. 803-9. Washington, D.C.: American Society for Microbiology 10.1128/JCM.02169-06

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Restriction fragment length polymorphism (RFLP) analysis is an economic and fast technique for molecular typing but has the drawback of difficulties in accurately sizing DNA fragments and comparing banding patterns on agarose gels. We aimed to improve RFLP for typing of the important human pathogen Streptococcus pneumoniae and to compare the results with the commonly used typing techniques of pulsed-field gel electrophoresis and multilocus sequence typing. We designed primers to amplify a noncoding region adjacent to the pneumolysin gene. The PCR product was digested separately with six restriction endonucleases, and the DNA fragments were analyzed using an Agilent 2100 bioanalyzer for accurate sizing. The combined RFLP results for all enzymes allowed us to assign each of the 47 clinical isolates of S. pneumoniae tested to one of 33 RFLP types. RFLP analyzed using the bioanalyzer allowed discrimination between strains similar to that obtained by the more commonly used techniques of pulsed-field gel electrophoresis, which discriminated between 34 types, and multilocus sequence typing, which discriminated between 35 types, but more quickly and with less expense. RFLP of a noncoding region using the Agilent 2100 bioanalyzer could be a useful addition to the molecular typing techniques in current use for S. pneumoniae, especially as a first screen of a local population.

Item Type:	Journal Article (Original Article)
Division/Institute:	04 Faculty of Medicine > Department of General Internal Medicine (DAIM) > Clinic of General Internal Medicine > Centre of Competence for General Internal Medicine 04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases
UniBE Contributor:	Brugger, Silvio, Mühlemann, Kathrin
ISSN:	0095-1137
ISBN:	17202282
Publisher:	American Society for Microbiology
Language:	English
Submitter:	Factscience Import
Date Deposited:	04 Oct 2013 14:56
Last Modified:	05 Dec 2022 14:17
Publisher DOI:	10.1128/JCM.02169-06
PubMed ID:	17202282
Web of Science ID:	000245071500016
URI:	https://boris.unibe.ch/id/eprint/23940 (FactScience: 45287)