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Open Access Highly Accessed Research article

Deletion of pic results in decreased virulence for a clinical isolate of Shigella flexneri 2a from China

Junqi Zhang12, Lisheng Qian1, Yang Wu2, Xia Cai2, Xueping Li1, Xunjia Cheng1* and Di Qu2*

Author affiliations

1 Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, No. 138 Yixueyuan Road, Shanghai, 200032, China

2 Key Laboratory of Medical Molecular Virology of Ministries of Education and Health, Institute of Medical Microbiology and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Fudan University, No. 138 Yixueyuan Road, Shanghai, 200032, China

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Citation and License

BMC Microbiology 2013, 13:31  doi:10.1186/1471-2180-13-31

Published: 8 February 2013

Abstract

Background

Shigella is a major pathogen responsible for bacillary dysentery, a severe form of shigellosis. Severity of the disease depends on the virulence of the infecting strain. Shigella pathogenicity is a multi-gene phenomenon, involving the participation of genes on an unstable large virulence plasmid and chromosomal pathogenicity islands.

Results

A multiplex PCR (mPCR) assay was developed to detect S. flexneri 2a from rural regions of Zhengding (Hebei Province, China). We isolated and tested 86 strains using our mPCR assay, which targeted the ipaH, ial and set1B genes. A clinical strain of S. flexneri 2a 51 (SF51) containing ipaH and ial, but lacking set1B was found. The virulence of this strain was found to be markedly decreased. Further testing showed that the SF51 strain lacked pic. To investigate the role of pic in S. flexneri 2a infections, a pic knockout mutant (SF301-pic) and two complementation strains, SF301-pic/pPic and SF51/pPic, were created. Differences in virulence for SF51, SF301-pic, SF301-pic/pPic, SF51/pPic and S. flexneri 2a 301 (SF301) were compared. Compared with SF301, both SF51 and SF301-pic exhibited lower levels of Hela cell invasion and resulted in reduced keratoconjunctivitis, with low levels of tissue damage seen in murine eye sections. The virulence of SF301-pic and SF51 was partially recovered in vitro and in vivo through the addition of a complementary pic gene.

Conclusions

The pic gene appears to be involved in an increase in pathogenicity of S. flexneri 2a. This gene assists with bacterial invasion into host cells and alters inflammatory reactions.

Keywords:
Shigella flexneri; Multiplex PCR; Clinical isolates; Pic gene; HeLa cell gentamicin protection assay; Mouse sereny tests