Restriction Fragment End Labeling Analysis: High-Resolution Genomic Typing of Streptococcus pneumoniae Isolates

2003 ◽  
pp. 169-179
Author(s):  
Peter W. M. Hermans ◽  
Marcel Sluijter ◽  
Alex van Belkum
Keyword(s):  
Streptococcus Pneumoniae ◽  
High Resolution ◽  
Restriction Fragment ◽  
Genomic Typing

scholarly journals A Novel Multiresistant Streptococcus pneumoniae Serogroup 19 Clone from Washington State Identified by Pulsed-Field Gel Electrophoresis and Restriction Fragment Length Patterns

2000 ◽  
Vol 38 (4) ◽  
pp. 1575-1580 ◽  
Author(s):  
Vicki A. Luna ◽  
Daniel B. Jernigan ◽  
Alan Tice ◽  
James D. Kellner ◽  
Marilyn C. Roberts
Keyword(s):  
Streptococcus Pneumoniae ◽  
Restriction Fragment ◽  
Fragment Length ◽  
Gel Electrophoresis ◽  
Washington State ◽  
Pulsed Field Gel Electrophoresis ◽  
Erythromycin Resistance ◽  
Eastern Canada ◽  
Pulsed Field ◽  
Restriction Fragment Length

In 1997, a cluster of multiresistant invasive serogroup 19 pneumococcus infections, including two fatalities, was reported in Washington State. Further investigation identified other cases. Fourteen Washington Streptococcus pneumoniae isolates, four from Alaska, and eight isolates from eastern Canada with reduced penicillin susceptibility (MIC of ≥1 μg/ml) were included in the study. Pulsed-field gel electrophoresis (PFGE) with ApaI,SacII, and SmaI restriction enzymes and IS1167 and mef restriction fragment length polymorphism (RFLP) pattern analysis were performed. Twenty of the 26 isolates had identical or related PFGE patterns, with two or all three enzymes, and identical or related IS1167 RFLP patterns, indicating that they were genetically related. These 20 isolates contained the mef gene conferring erythromycin resistance and had identical mef RFLP patterns. The PFGE and RFLP patterns were distinct from those of six multiresistant clones previously described and suggest that a new multiresistant clone has appeared in Washington, Alaska, and eastern Canada. This newly characterized clone should be included in the Pneumococcal Molecular Epidemiology Network.

scholarly journals Identification of novel non-coding small RNAs from Streptococcus pneumoniae TIGR4 using high-resolution genome tiling arrays

BMC Genomics ◽  
2010 ◽  
Vol 11 (1) ◽  
pp. 350 ◽  
Author(s):  
Ranjit Kumar ◽  
Pratik Shah ◽  
Edwin Swiatlo ◽  
Shane C Burgess ◽  
Mark L Lawrence ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
High Resolution ◽  
Small Rnas ◽  
Tiling Arrays

scholarly journals Novel BOX repeat PCR assay for high-resolution typing of Streptococcus pneumoniae strains.

1996 ◽  
Vol 34 (5) ◽  
pp. 1176-1179 ◽  
Author(s):  
A van Belkum ◽  
M Sluijuter ◽  
R de Groot ◽  
H Verbrugh ◽  
P W Hermans
Keyword(s):  
Streptococcus Pneumoniae ◽  
High Resolution ◽  
Pcr Assay

scholarly journals Curvas de fusión de regiones genómicas específicas: una herramienta prometedora para el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia

Biomédica ◽  
2017 ◽  
Vol 37 (4) ◽  
pp. 538 ◽  
Author(s):  
Johana Marin ◽  
Daniel Urrea ◽  
Carlos Muskus ◽  
María Clara Echeverry ◽  
Ana María Mejía ◽  
...  
Keyword(s):  
High Resolution ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
High Resolution Melting ◽  
Fragment Length Polymorphism ◽  
Pcr Rflp ◽  
Leishmania Panamensis ◽  
Restriction Fragment Length

Introducción. La leishmaniasis cutánea es una enfermedad causada por parásitos del género Leishmania que tiene gran incidencia en Colombia. El diagnóstico y la identificación de la especie infecciosa son factores críticos en el momento de escoger e iniciar el tratamiento. Actualmente, los métodos de diagnóstico y tipificación requieren procedimientos complejos, por lo que es necesario validar nuevos marcadores moleculares y métodos que simplifiquen el proceso.Objetivo. Desarrollar una herramienta basada en la reacción en cadena de la polimerasa (PCR) con curvas de fusión (High Resolution Melting; PCR-HRM) para el diagnóstico y tipificación de las tres especies de Leishmania de importancia epidemiológica en casos de leishmaniasis cutánea en Colombia.Materiales y métodos. Los genomas de Leishmania panamensis, L. braziliensis y L. guyanensis se compararon mediante métodos bioinformáticos. Las regiones específicas de especie identificadas se validaron mediante PCR. Para los marcadores seleccionados se diseñó una PCR-HRM y se estimaron algunos parámetros de validez y seguridad usando aislamientos de pacientes colombianos caracterizados previamente mediante PCR y análisis de polimorfismos en la longitud de los fragmentos de restricción (Restriction Fragment Length Polymorphism – RFLP; PCR-RFLP) del gen hsp70.Resultados. El análisis genómico comparativo mostró 24 regiones específicas de especie. Sin embargo, la validación mediante PCR solo identificó un marcador específico para cada especie de Leishmania. Los otros marcadores mostraron amplificación cruzada. El límite de detección para los tres marcadores seleccionados fue de un parásito, mientras que la sensibilidad, la especificidad, el valor predictivo positivo y el negativo fueron de 91,4, 100, 100 y 75 %, respectivamente.Conclusiones. Las tres regiones seleccionadas pueden emplearse como marcadores moleculares en el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia.

High-resolution NMR spectroscopic analysis of the C-polysaccharide of Streptococcus pneumoniae

1993 ◽  
Vol 71 (5) ◽  
pp. 644-648 ◽  
Author(s):  
Malgorzata Kulakowska ◽  
Jean-Robert Brisson ◽  
Douglas W. Griffith ◽  
N. Martin Young ◽  
Harold J. Jennings
Keyword(s):  
Cell Wall ◽  
Streptococcus Pneumoniae ◽  
Nmr Spectroscopy ◽  
High Resolution ◽  
Spectroscopic Analysis ◽  
Theoretical Calculations ◽  
Complex Cell ◽  
Complete Structure ◽  
High Resolution Nmr

The complete structure of the complex cell-wall C-polysaccharide from Streptococcus pneumoniae R36A was elucidated using a combination of high-resolution NMR spectroscopy and theoretical calculations. The C-polysaccharide is composed of the following repeating unit β-D-Glcp1 → 3-α-D-AATp1 → 4-α-D-GalpNAc1 → 3-β-D-GalpNAc1 → 1-ribitol-5-phosphate where AAT is 2-acetamido-4-amino-2,4,6-trideoxygalactopyranoside. The C-polysaccharide contains two phosphorylcholine substituents per repeating unit, which are situated on O-6 of each of the two 2-acetamido-2-deoxy-galactopyranose residues. The repeating units are linked through a diphosphate ester from ribitol to O-6 of the β-D-glucopyranosyl residue.

Structure of the specific capsular polysaccharide of Streptococcus pneumoniae type 23F (American type 23)

1988 ◽  
Vol 66 (7) ◽  
pp. 758-771 ◽  
Author(s):  
James C. Richards ◽  
Malcolm B. Perry
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Streptococcus Pneumoniae ◽  
High Resolution ◽  
Capsular Polysaccharide ◽  
Optical Rotation ◽  
Periodate Oxidation ◽  
Partial Hydrolysis ◽  
Composition Analysis ◽  
Part D ◽  
Magnetic Resonance Studies

The specific capsular polysaccharide of Streptococcus pneumoniae serotype 23F (American type 23) is composed of a repeating tetrasaccharide unit containing D-glucose (one part), D-galactose (one part), L-rhamnose (two parts), glycerol (one part), and phosphate (one part). By composition analysis, optical rotation, partial hydrolysis, periodate oxidation, methylation, and high-resolution 1H and 13C nuclear magnetic resonance studies, the elucidated unambiguous structure was in agreement with our earlier proposal but is at variance with structures proposed later by other authors. The structure of the type 23F pneumococcal polysaccharide is[Formula: see text]

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