Epidemiology and evolution of genotype and antimicrobial resistance of an imported Shigella sonnei clone circulating in central Taiwan

Wei, Hsiao-Lun; Wang, Youwen; Li, Chun-Chin; Tung, Shu-Chu; Chiou, Chien-Shun

doi:10.1016/j.diagmicrobio.2007.03.006

Cited by 37 publications

(38 citation statements)

References 16 publications

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“…It caused a large outbreak of shigellosis at a high school in October 2000, and thereafter it and its derivative strains circulated throughout the country for 3.25 years (15). Since these strains were derived from a line of common evolutionary descent, they were an excellent collection of isolates to identify the highly variable VNTR loci and investigate the phylogenetic pattern for the newly emerged clone by use of combinations of loci with different variation values.…”

Section: Discussionmentioning

confidence: 99%

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Utility of Multilocus Variable-Number Tandem-Repeat Analysis as a Molecular Tool for Phylogenetic Analysis ofShigella sonnei

Chiou

Watanabe

Wang³

et al. 2009

J Clin Microbiol

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A panel of 916 isolates, including 703 closely related IST1 isolates, were characterized by inter-IS1 spacer typing (IST), pulsed-field gel electrophoresis (PFGE), and multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) to evaluate the utility of MLVA as a molecular tool for the phylogenetic analysis of Shigella sonnei. The global phylogenetic patterns determined by IST, PFGE, and MLVA were concordant. MLVA was carried out using 26 VNTR loci with a range of degrees of variability. MLVA data for the 703 IST1 isolates revealed that diversification among the closely related isolates was attributed mainly to four highly variable loci. The phylogenetic pattern for the closely related isolates determined using MLVA profiles of 8 highly variable loci was in agreement with that determined using the 26-locus profiles. A clustering analysis using the profiles of 18 loci with limited variability established clear phylogenetic relationships among IST clonal groups. Accordingly, MLVA is a useful tool for the phylogenetic analysis of S. sonnei. Combined VNTR loci with higher variability are useful markers for resolving closely related isolates, whereas combined loci with lower variability are suitable for establishing clear phylogenetic relationships between strains or clones that have evolved over a longer timescale.Shigella sonnei is one of the causative agents of shigellosis. Unlike the other three Shigella species, S. dysenteriae, S. flexneri, and S. boydii, which are prevalent in developing countries, S. sonnei is predominant in industrialized countries (6) and is one of the major causes of travel-associated diarrheal disease (3). Transmission of S. sonnei strains between countries occurs frequently via international travel (7,14).The analysis of bacterial isolates by various genotyping methods provides useful information for establishing the genetic relatedness among isolates for the purposes of epidemiological investigation and phylogenetic study. Among these genotyping methods, pulsed-field gel electrophoresis (PFGE) has been proven to be a powerful tool for discriminating Shigella strains and has become a standardized method for an international molecular-subtyping network for food-borne-disease surveillance (13). However, PFGE is, at times, too discriminatory for investigating clonal relationships among Shigella strains that have evolved over years or decades. In order to study an S. sonnei epidemic, we previously developed an inter-IS1 spacer typing (IST) method for subtyping of S. sonnei strains (1). IST is less discriminative than PFGE for S. sonnei, but it is more suitable than PFGE for investigating the clonal relationships among S. sonnei strains in circulation over a short timescale (15).Multilocus sequence typing (MLST) is a sequence-based typing method widely adopted for the phylogenetic study of a number of bacterial pathogens, as listed on the website http: //www.mlst.net/. The MLST method has been used for phylogenetic analysis of Shigella spp. with the protocol developed for Escherichia coli...

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Section: Discussionmentioning

confidence: 99%

“…In order to study an S. sonnei epidemic, we previously developed an inter-IS1 spacer typing (IST) method for subtyping of S. sonnei strains (1). IST is less discriminative than PFGE for S. sonnei, but it is more suitable than PFGE for investigating the clonal relationships among S. sonnei strains in circulation over a short timescale (15).…”

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confidence: 99%

Utility of Multilocus Variable-Number Tandem-Repeat Analysis as a Molecular Tool for Phylogenetic Analysis ofShigella sonnei

Chiou

Watanabe

Wang³

et al. 2009

J Clin Microbiol

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“…S. flexneri es conocida por ser predominante en países en vías de desarrollo y S. sonnei en países industrializados [1,4]. Estudios recientes han indicado que S. sonnei está superando a las poblaciones de S. flexneri en países como Tailandia, Korea del Sur y Taiwan [17,18]. En la presente investigación se observó que la especie más común resultó ser S. flexneri, seguido por S. sonnei y S. boydii y finalmente no se reportó presencia de S. dysenteriae.…”

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Sensibilidad antimicrobiana entre los serogrupos de <em>Shigellae</em> aislados en la ciudad de Quito-Ecuador

Granda

Alcocer

2015

Av. Cienc. Ing. (Quito)

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The pathotype Shigellae includes bacillus from the species S. flexneri, S. sonnei, S. boydii and S. dysenteriae which are causatives of the disease known as bacillary dysentery or shigellosis. The pathogenicity of the genus is characterized by the presence of high antibiotic resistance. The aim of this study was to identify the species and antimicrobial susceptibility in clinical isolates of Shigellae. We analyzed 79 isolates obtained from Zurita & Zurita Laboratorios and Vozandes Hospital in the city of Quito. Three species were obtained by serotyping, using polyclonal antisera: S. flexneri (n=50; 63,29 %), S. sonnei (n=23; 29,11 %), and S. boydii (n=6; 7,59 %). The antimicrobial susceptibility was analyzed using the Kirby Bauer method and recommendations from the "Clinical and Laboratory Standards Institute" (CLSI). Most of the isolates showed resistance to tetracycline (n=76; 96,20 %), ampicillin (n=75; 94,94 %), trimethoprim/sulfamethoxazole (n= 68; 86,08 %) and chloramphenicol (n=67; 84,81 %). The results showed the incidence of multidrug resistance to antibiotics commonly used for treating shigellosis, and the presence of three species of Shigellae.Keywords. Antibiotics, bacillary dysentery, Shigellae. ResumenEl patotipo Shigellae comprende bacilos de las especies S. flexneri, S. sonnei, S. boydii y S. dysenteriae, causantes de la enfermedad conocida como disentería bacilar o shigelosis. La patogenicidad del género se caracteriza por la presencia de una alta multirresistencia a antibióticos. El objetivo de este estudio fue identificar las especies y la sensibilidad a antimicrobianos en aislados clínicos de Shigellae. Se analizaron 79 aislados obtenidos de Zurita & Zurita Laboratorios y del Hospital Vozandes en la ciudad de Quito. Mediante serotipaje, con el uso de antisueros policlonales, se obtuvieron 3 especies: S. flexneri (n=50; 63,29 %), S. sonnei (n= 23; 29,11 %), y S. boydii (n= 6; 7,59 %). La sensibilidad antimicrobiana se analizó siguiendo el método de Kirby-Bauer y las recomendaciones del "Clinical and Laboratory Standards Institute" (CLSI). La mayoría de las cepas mostraron resistencia a tetraciclina (n=76; 96,20 %), ampicilina (n=75; 94,94 %), trimetoprim/sulfametoxazol (n=68; 86,08 %) y cloranfenicol (n=67; 84,81 %). Los resultados obtenidos demostraron la incidencia de multirresistencia a antibióticos comúnmente utilizados para el tratamiento de shigelosis, y la presencia de 3 especies de Shigellae.Palabras Clave. Antibióticos, disentería bacilar, Shigellae. IntroducciónEl patotipo Shigellae comprende bacilos Gram negativos de 0,3 a 1 µm diámetro y de 1 a 6 µm de longitud, no móviles, no formadores de esporas, anaerobios facultativos, pertenecientes a la familia Enterobacteriaceae y causantes de la enfermedad conocida como shigelosis o disentería bacilar [1]. Su identificación se fundamenta en características bioquímicas y antigénicas, en base a ellas se describen cuatro especies: Shigella dysenteriae, serogrupo A; Shigella flexneri, serogrupo B; Shigella boydii, serogrupo C; y, Shige...

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“…higella sonnei, an emerging global cause of shigellosis, consists of four distinct lineages, and the current pandemic involves several geographically associated, multidrug-resistant clones that belong to lineage III (1)(2)(3). A typing scheme based on high-resolution melting (HRM) of six chromosomal single nucleotide polymorphisms (SNPs) has been described to identify all lineages/ sublineages (4).…”

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Rapid Genotyping of Shigella sonnei by Use of Multiplex High-Resolution Melting

et al. 2015

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Shigella sonnei, an emerging global cause of shigellosis, consists of four distinct lineages, and the current pandemic involves several geographically associated, multidrug-resistant clones that belong to lineage III (1-3). A typing scheme based on high-resolution melting (HRM) of six chromosomal single nucleotide polymorphisms (SNPs) has been described to identify all lineages/ sublineages (4). HRM detects SNPs in small DNA fragments based on the melting patterns (5, 6). HRM involves minimal labor, with a rapid turnaround time and a decreased risk of PCR carryover contamination (7). A sensitive intercalating fluorescent dye, EvaGreen, may yield robust fluorescence intensities and strong sharp melting peaks upon melting of the PCR amplicons (8-10). Based on the SNPs summarized in Table 1, we have developed a multiplex HRM assay for identification of lineages I, II, and III using the EvaGreen dye and a Rotor-Gene 6000 instrument (Qiagen).Ten strains (2 for each lineage/sublineage) were from a collection held at the Institut Pasteur. Only one lineage IV strain is reported so far (1), which is not included in the study. Primers were designed to amplify fragments that contain three SNPs within kduD, deoA, and emrA for typing 3 main lineages in a first set run and to amplify fragments that contain SNPs within fdX and menF for typing sublineages IIIa, IIIb, and IIIc in a second run. Primer sequences are summarized in Table 2.In the first run, we set up a 20-l multiplex real-time PCR mixture containing 10 l SensiMix with EvaGreen dye (Bioline), 20 ng of template DNA, and 2.5 l of each primer of kduD (100 M), 1 l of each primer of deoA (100 M), and 1 l of each primer of emrA (50 M). In the second set run, the 20-l reaction mix was the same, except that the primers used were for fdX (10 M) and menF (10 M), with a quantity of 2 l and 1 l, respectively. The PCR was programmed as follows: the mixture was held at 95°C for 10 min and then subjected to 35 cycles of 95°C for 20 s, 60°C for 20 s, and 72°C for 22 s. HRM was performed between 80 and 90°C, with a stepwise increase of 0.05°C. The melting temperatures (T m ) were determined by the negative derivative of the decrease in fluorescence over the increase in temperature (df/dt) for each SNP using the proprietary software (version 1.7.34).The first set run clearly separated lineages I, II, and III with distinctive melting curves, and the T m of each allele was at least a half degree apart from that of the other alleles (Fig. 1A). The second set run distinguished the sublineages IIIa, IIIb, and IIIc with

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Epidemiology and evolution of genotype and antimicrobial resistance of an imported Shigella sonnei clone circulating in central Taiwan

Cited by 37 publications

References 16 publications

Utility of Multilocus Variable-Number Tandem-Repeat Analysis as a Molecular Tool for Phylogenetic Analysis ofShigella sonnei

Utility of Multilocus Variable-Number Tandem-Repeat Analysis as a Molecular Tool for Phylogenetic Analysis ofShigella sonnei

Sensibilidad antimicrobiana entre los serogrupos de <em>Shigellae</em> aislados en la ciudad de Quito-Ecuador

Rapid Genotyping of Shigella sonnei by Use of Multiplex High-Resolution Melting

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