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Shigellosis, caused by  Shigella boydii  type 1, is understudied and underreported. For 3 years, GEMS study identified 5.4% of all  Shigella  as  S. boydii . We showed the prevalent serotypes of  S. boydii  in Bangladesh and phage-based diagnosis of  S. boydii  type 1, a rapid and low-cost approach. Previously typed 793 clinical  S. boydii  strains were used for serotype distribution. Twenty-eight environmental water samples were collected for isolation of  Shigella  phages. Forty-eight serotypes of  Shigella  and other enteric bacteria were used for testing the susceptibility to phage MK-13. Electron microscopy, restriction enzyme analysis, whole genome sequencing (WGS), and annotation were performed for extensive characterization.  S. boydii  type 1 is the second most prevalent serotype among 20 serotypes of  S. boydii  in Bangladesh. We isolated a novel phage, MK-13, which specifically lyses  S. boydii  type 1, but doesn’t lyse other 47 serotypes of  Shigella  or other enteric bacteria tested. The phage belongs to the  Myoviridae  family and distinct from other phages indicated by electron microscopy and restriction enzyme analysis, respectively. MK-13 genome consists of 158 kbp of circularly permuted double-stranded DNA with G + C content of 49.45%, and encodes 211 open reading frames including four tRNA-coding regions. The genome has 98% identity with previously reported phage, ΦSboM-AG3, reported to have a broader host range infecting most of the  S. boydii  and other species of  Shigella  tested. To our knowledge, MK-13 is the first phage reported to be used as a diagnostic marker to detect  S. boydii  type 1, especially in remote settings with limited laboratory infrastructure.

frontiers in microbiology

Prevalence of Shigella boydii in Bangladesh: Isolation and Characterization of a Rare Phage MK-13 That Can Robustly Identify Shigellosis Caused by Shigella boydii Type 1