Molecular characterization of serologically atypical provisional serovars of Shigella isolates from Kolkata, India

During 2000-2004, 13 Shigella strains that were untypable by commercially available antisera were isolated from children <5 years of age with acute diarrhoea in Kolkata. These strains were subsequently identified as Shigella dysenteriae provisional serovar 204/96 (n = 3), Shigella dysenteriae provisional serovar E23507 (n = 1), Shigella dysenteriae provisional serovar I9809-73 (n = 1), Shigella dysenteriae provisional serovar 93-119 (n = 1), Shigella flexneri provisional serovar 88-893 (n = 6) and Shigella boydii provisional serovar E16553 (n = 1). In this study, characterization of those provisional serovars of Shigella was performed with respect to their antimicrobial resistance, plasmids, virulence genes and PFGE profiles. The drug resistant strains (n = 10) of Shigella identified in this study possessed various antibiotic resistance genetic markers like catA (for chloramphenicol resistance); tetA and tetB (for tetracycline resistance); dfrA1 and sul2 (for co-trimoxazole resistance); aadA1, strA and strB (for streptomycin resistance) and blaOXA-1 (for ampicillin resistance). Class 1 and/or class 2 integrons were present in eight resistant strains. Three study strains were pan-susceptible. A single mutation in the gyrA gene (serine to leucine at codon 83) was present in four quinolone resistant strains. The virulence gene ipaH (invasion plasmid antigen H) was uniformly present in all strains in this study, but the stx (Shiga toxin) and set1 (Shigella enterotoxin 1) genes were absent. Other virulence genes like ial (invasion associated locus) and sen (Shigella enterotoxin 2) were occasionally present. A large plasmid of 212 kb and of incompatibility type IncFIIA was present in the majority of the strains (n = 10) and diversity was noticed in the smaller plasmid profiles of these strains even within the same provisional serovars. PFGE profile analysis showed the presence of multiple unrelated clones among the isolates of provisional Shigella serovars. To the best of our knowledge, this is the first report on the phenotypic and molecular characterization of provisional serovars of Shigella isolates from Kolkata, India.