<sec><title>Objectives</title><p>The genus <italic>Shigella</italic> comprises the most infectious and diarrheagenic bacteria causing severe diseases, mostly in children under five years of age. This study aimed to detect nine virulence genes (<italic>ipaBCD</italic>, <italic>VirA</italic>, <italic>sen</italic>, <italic>set1A</italic>, <italic>set1B</italic>, <italic>ial</italic>, <italic>ipaH</italic>, <italic>stx</italic>, and <italic>sat</italic>) in <italic>Shigella</italic> species (spp.) using multiplex polymerase chain reaction (MPCR) and to determine the relation of <italic>Shigella</italic> spp. from pediatric diarrheal samples with hospitalization and bloody diarrhea in Tehran, Iran.</p></sec><sec><title>Methods</title><p><italic>Shigella</italic> spp. were isolated and identified using standard microbiological and serological methods. The virulence genes were detected using MPCR.</p></sec><sec><title>Results</title><p>Seventy-five <italic>Shigella</italic> spp. (40 <italic>S. sonnei</italic>, 33 <italic>S. flexneri</italic>, 1 <italic>S. dysenteriae</italic>, and 1 <italic>S. boydii</italic>) were isolated in this study. The prevalence of <italic>ial</italic>, <italic>sen</italic>, <italic>sat</italic>, <italic>set1A</italic>, and <italic>set1B</italic> was 74.7%, 45.4%, 28%, 24%, and 24%, respectively. All <italic>S. flexneri</italic> isolates, while no <italic>S. sonnei</italic>, <italic>S. dysenteriae</italic>, or <italic>S. boydii</italic> isolates, contained <italic>sat</italic>, <italic>set1A</italic>, and <italic>set1B</italic>. All isolates were positive for <italic>ipaH</italic>, <italic>ipaBCD</italic>, and <italic>virA</italic>, while one (1.4%) of the isolates contained <italic>stx</italic>. The highest prevalence of virulence determinants was found in <italic>S. flexneri</italic> serotype IIa. Nineteen (57.6%) of 33 <italic>S. flexneri</italic> isolates were positive for <italic>ipaBCD</italic>, <italic>ipaH</italic>, <italic>virA</italic>, <italic>ial</italic>, and <italic>sat</italic>. The <italic>sen</italic> determinants were found to be statistically significantly associated with hospitalization and bloody diarrhea (<italic>p</italic> = 0.001).</p></sec><sec><title>Conclusion</title><p>This study revealed a high prevalence of enterotoxin genes in <italic>S. flexneri</italic>, especially in serotype 2a, and has presented relations between a few clinical features of shigellosis and numerous virulence determinants of clinical isolates of <italic>Shigella</italic> spp.</p></sec>
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