Abstract
Background: Shigella is one of the most serious pathogens associated with bloody diarrhea in children. The empiric antibiotic therapy of enteric illness with blood streaked stool leads to emergence of multi drug resistant (MDR) Shigella. The condition gets exacerbated by presence of integrons that facilitate the horizontal spread. Virulence genes associated with MDR Shigella modulate the patient outcome, particularly in children. Objectives: The present study was aiming at isolation of MDR Shigella from children with diarrheal sickness and characterization of those isolates as regarding presence of class 1 integrase and other virulence genes. Methods: Four hundred and ninety patients under the age of five suffering from diarrheal illness were examined for presence of Shigella in their stool specimens. MDR Shigella was determined using the antibiotic susceptibility testing by disc diffusion method; those isolates were tested for presence of class 1 integrase by PCR. Multiplex PCR assay was used to determine the presence of virulence genes, virA, ial, sen, set1A, set1B, sat, ipaBCD, ipaH and stx in the MDR Shigella isolates. Results: The isolation rate of Shigella from pediatric patients was 5.3%. Most of the isolated Shigella (57.7%) were from infants between 12 and 23 month. 73.1% of the identified Shigella were MDR. intI1 gene was present in 78.9% of MDR isolates. Muliplex PCR revealed that ipaH and ipaBCD, virA, sat, ial, set1A and set1B, sen were detected in 94.7%, 78.9%, 73.7%, 68.4%, 42.1%, 36.8% of the MDR Shigella isolates respectively. Conclusion: The MDR isolates represented a considerable percentage of Shigella detected in pediatric patients. Presence of intI1 gene in most of MDR Shigella reflects the higher possibility of resistant strains spread. Existence of a variety of virulence genes in those isolates is an important indicator of serious disease outcome.
Highlights
Shigella is one of the most important pathogens associated with a serious food born diarrheal disease termed shigellosis [1]
Diarrheal diseases caused by Shigella usually require antibiotic administration to decrease the period of the disease and limit its spread among close associates; resistance to trimethoprim-sulfamethoxazole and ampicillin have been continuously emerged by Shigella isolates due to their excessive usage; ceftriaxone and flouroquinolones are suggested by the WHO for management of dysentery caused by Shigella in pediatric patients [8], and they are efficient against Shigella isolates exhibiting multidrug resistance in immunosuppressed children [9]
Subculture was done on Salmonella Shigella (SS) agar, Xylose Lysin Desoxycholate (XLD) agar and Hekton Enteric (HE) agar media, Shigella isolates appeared as non lactose fermenting colonies on the plates were selected for further identification by the standard biochemical reactions and the API 20E (Bio-merieux SA, Montalieu Vercica and France) Antibiotic susceptibility of Shigella isolates: The disk diffusion technique was used to assess the antibiotic sensitivity for all isolated Shigella according to the CLSI, 2014 [25]
Summary
Shigella is one of the most important pathogens associated with a serious food born diarrheal disease termed shigellosis [1]. Diarrhea streaked with blood in pediatrics is seriously considered as an important manifestation of enteric disease associated with invasion which usually leads to high morbidity and mortality outcomes; in developing countries, the commonest bacteria that mostly recovered from fecal samples of children suffering from blood streaked diarrhea are Shigella [7]. Objectives: The present study was aiming at isolation of MDR Shigella from children with diarrheal sickness and characterization of those isolates as regarding presence of class 1 integrase and other virulence genes. Multiplex PCR assay was used to determine the presence of virulence genes, virA, ial, sen, set1A, set1B, sat, ipaBCD, ipaH and stx in the MDR Shigella isolates. Existence of a variety of virulence genes in those isolates is an important indicator of serious disease outcome
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