Polymorphisms in the genes encoding surface associated proteins of Clostridioides difficile isolates

Background: Although the diversity of Clostridioides difficile toxins have been extensively studied, little is known about the variation in the surface associated proteins (SAPs) which are important in early steps of bacterial colonization and infection. Here, we examined 65C. difficile isolates to identify polymorphisms in the genes encoding SAPs. Methods: PCR was used to amplify slpA, fliC, fliD, cwp66 and cwp84 genes, followed by sequencing. In addition, the antigenicity and immunogenicity properties of different types of SlpA, FliC, FliD, Cwp66 and Cwp84 proteins were predicted in-silico by VaxiJen and BcePred online servers. Results: The predominant slpA sequence type was gr-01 (42.37), followed by hr-01 (11.86) and 078-01 (10.16). In addition, two new slpA subtypes of smz (smz-09-Ir and smz-010-Ir) and a new slpA sequence type (Ir-01) were identified among the isolates examined. Analysis of the nucleotide sequences of fliC, fliD, cwp66 and cwp84 genes revealed 7, 5,5,3 different sequence types, respectively. Insilico analysis of antigenicity of SAPs showed that FliC had the highest level of antigenicity whereas SlpA and Cwp66 proteins had the highest level of immunogenicity. Conclusions: This study pointed to the nucleotide polymorphism in SAPs of C. difficile isolates and demonstrated noticeable diversity in antigenicity and immunogenicity of these proteins which need to be taken into consideration as promising therapeutic or vaccine targets. © 2020 Elsevier B.V