Molecular Genotyping of Shigella sonnei Strains Isolated From Children With Bloody Diarrhea Using Pulsed Field Gel Electrophoresis on the Total Genome and PCR-RFLP of IpaH and IpaBCD Genes

Farshad, Shohreh; Ranjbar, Reza; Hosseini, Mona

doi:10.5812/jjm.14004

Cited by 15 publications

(21 citation statements)

References 30 publications

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“…The ipaH gene is present in numerous copies in the chromosome and in the virulence plasmid of species of the genus Shigella, and is associated with the invasion of bacteria into host cells (Cruz et al, 2014;Farshad et al, 2015). The ipaBCD and ial plasmidial genes are responsible for the invasion of species of the genus Shigella (Campos et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Molecular and phenotypic characterization of strains of Shigella sonnei isolated over 31 years suggests the circulation of two prevalent subtypes in São Paulo State, Brazil

Seribelli

Frazão

Medeiros

et al. 2016

Journal of Medical Microbiology

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Shigella sonnei is an important causative agent of bacillary dysentery worldwide that has recently emerged in developing countries. However, there are few studies that have characterized strains ofS. sonnei isolated in Brazil. The aims of this study were to assess the presence of 12 virulence genes, the antimicrobial resistance profile against 16 drugs and the genotypic diversity of strains of S. sonnei isolated in this country. Seventy-two strains of S. sonnei isolated from human diarrhoeic faeces in São Paulo State, Brazil from 1983-2014 were studied. All of the strains contained the ipaH, iuc and sigA genes. The ipaBCD gene was detected in 19 % of the strains, the ial and virF genes in 18 % and the sen gene in 10 % of the strains. The set1A, set1B, pic, sepA and sat genes were not detected. A total of 42 (58.3 %) strains were resistant to trimethoprim-sulfamethoxazole. Thirty (41.6 %) strains were resistant to tetracycline. The S. sonnei strains were grouped in two clusters called A and B by PFGE and ERIC-PCR, and the majority of the strains comprised in each cluster presented !80 % similarity. In conclusion, the pathogenic potential of the strains studied was highlighted by the presence of important virulence genes. The high rates of resistance to trimethoprim-sulfamethoxazole and tetracycline are alarming once those drugs can be used in the treatment of shigellosis. The PFGE and ERIC-PCR results suggest that there are two prevalent subtypes in the studied strains of S. sonnei that differed little over 31 years and have been contaminating humans and causing diseases in São Paulo State, Brazil.

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Section: Introductionmentioning

confidence: 99%

Molecular and phenotypic characterization of strains of Shigella sonnei isolated over 31 years suggests the circulation of two prevalent subtypes in São Paulo State, Brazil

Seribelli

Frazão

Medeiros

et al. 2016

Journal of Medical Microbiology

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“…Ampicillin and sulfamethoxazole-trimethoprim are commonly used as the first-line drugs for the therapy of shigellosis while second-line drugs including the third generation of cephlosporines and fluoroquinolones are used for the treatment of children and adults respectively (11,16). Unluckily, it has been recently reported that the prevalence of resistance to some commonly used antibiotics is increasing in Shigella spp in Iran (17,18 (5,11,19). In our study, the prevalence of ESBL producing Shigella isolates accounted for 11.4 % of all Shigella isolates which is larger than the reports from Iran and other countries (11,20,21).…”

Section: Discussionmentioning

confidence: 99%

“…S. flexneri is the most common agent of shigellosis in developing countries and tropical areas (e.g., Iran). However, epidemiological shift in shigellosis and the change in the pattern of antibiotic resistance among Shigella strains have been reported in recent years in Iran and the world (5,6). As with other infectious diarrheal diseases, treatment of shigellosis includes both rehydration and antibiotics.…”

Section: Introductionmentioning

confidence: 99%

“…Plasmid profile analysis has been used as an epidemiological tool for investigating the outbreak of infectious diseases. Plasmid profile analysis may be useful in identifying the source of infection, discriminating strains or assessing the ability of control procedure (5,8). So far, plasmid profile analysis for typing Shigella strains in Ahvaz has not been evaluated.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of Shigella Strains by Plasmid Profile Analysis and Antibiotic Susceptibility Patterns in a Pediatric Hospital in Ahvaz

Sakhaei¹,

Savari²,

Shokoohizadeh³

et al. 2015

Int J Enteric Pathog

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Background: High incidences of dysentery and diarrhea were reported in a pediatric hospital in Ahvaz, Iran during March to April, 2013. Objectives: A cross-sectional study was therefore undertaken to identify the causative agents. Patients and Methods: A total of 230 diarrhea samples were collected from the patients and analyzed by routine bacteriological methods. Bacterial identification, serological assay, antimicrobial susceptibility testing, extended spectrum β-lactamases (ESBLs) screening and plasmid profile analysis were performed according to the standard guidelines. Results: A total of 70 Shigella strains including %70 (n = 49) S. sonnei and 30% (n = 21) S. flexneri were isolated from diarrhea samples. Most of the Shigella isolates showed high degrees of resistance to ampicillin, ulafamethoxazole-trimethoprime and cefexim. Concurrent resistance to sulafametoxazole-trimethoprime and ampicillin was the most common resistance pattern. Overall, 11.4% of Shigella isolates showed the ESBL producer criteria. The plasmid profile patterns of all the strains were determined by a modified alkaline lysis method. By plasmid profile analysis 23 genotypes were identified among all the isolates, 14 and 9 genotypes among the S. sonnei and S. flexneri respectively. S. sonnei and S. flexneri isolates demonstrated unique plasmid profiles. Conclusions: These data demonstrated that S. sonnei strains are the main cause of shigellosis as the prevalent Shigella serotype in Iran. We also found that the antibiotic resistance rates are increasing among Shigella strains. Plasmid profile analysis is more reliable than antibiotic susceptibility patterns in epidemiologic studies.

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“…Considering the presence of these repetitive extragenic palindromic sequences, by designing an appropriate primer according to desired sequence using PCR, electrophoresis of the resultant PCR products, and gel analysis by computer (6)(7)(8)(9) and given the number of sequences in each strain and their difference in length, the number of bonds obtained by PCR for each strain is different, and therefore the strains are classified based on diversity of bonds. Although, use of PFGE is considered as a gold standard method, the REP-PCR is a simpler and lower-cost method and provides more rapid results (11).…”

Section: Introductionmentioning

confidence: 99%

Molecular Analysis of Shigella sonnei Strains Isolated from Hospitals of Tehran and Ilam Cities Using Repetitive Extragenic Palindromic Polymerase Chain Reaction (REP-PCR)

et al. 2018

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Background and Objective: Shigella species belong to the family Enterobacteriaceae, which cause dysentery with abdominal pain and tenesmus in human. Our country is one of the endemic and occasional epidemic areas of the disease. In the present study, Repetitive Extragenic Palindromic Polymerase Chain Reaction (REP-PCR) technique that has high differentiation and specificity power compared to phenotypic markers, was used to investigate diversity and geographical distribution of colones as well as typing of Shigella strains.Methods: In this study, a total of 40 Shigella sonnei samples were isolated from stool of patients with diarrhea and divided into 5 groups. After identification and confirmation of the isolates by biochemical and serotyping methods, one colony of each isolate was cultured on LB medium and after DNA extraction, PCR was performed. After electrophoresis of the PCR product, gel images were saved in the computer for further analysis and typing of the isolates.Result: In this research, 40 isolates were examined using REP-PCR and a similarity matrix was constructed based on Dice's coefficient. According to this matrix, some isolates were completely similar (genetic similarity coefficient= 1) and some isolates showed the least similarity (genetic similarity coefficient= 0). The dendrogram was obtained using the UPGMA algorithm. The calculated Cophenetic correlation coefficient for this dendrogram was 0.91645. Conclusion:From the results of the present study, it was concluded that we can type the Shigella sonnei strains using palindromic repetitive sequences. The extent of polymorphism indicates that REP-PCR technique is a useful method for genetic variation analysis in molecular typing of shigella sonnei strains. Although, some strains were completely similar, a high genetic variation was found among the studied population of Shigella sonnei. This level of variation is probably due to wide geographic distribution of this species in Iran.

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Molecular Genotyping of Shigella sonnei Strains Isolated From Children With Bloody Diarrhea Using Pulsed Field Gel Electrophoresis on the Total Genome and PCR-RFLP of IpaH and IpaBCD Genes

Cited by 15 publications

References 30 publications

Molecular and phenotypic characterization of strains of Shigella sonnei isolated over 31 years suggests the circulation of two prevalent subtypes in São Paulo State, Brazil

Molecular and phenotypic characterization of strains of Shigella sonnei isolated over 31 years suggests the circulation of two prevalent subtypes in São Paulo State, Brazil

Characterization of Shigella Strains by Plasmid Profile Analysis and Antibiotic Susceptibility Patterns in a Pediatric Hospital in Ahvaz

Molecular Analysis of Shigella sonnei Strains Isolated from Hospitals of Tehran and Ilam Cities Using Repetitive Extragenic Palindromic Polymerase Chain Reaction (REP-PCR)

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