Updated campylobacter jejuni capsule PCR multiplex typing system and its application to clinical isolates from south and southeast Asia

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Abstract

Campylobacter jejuni produces a polysaccharide capsule that is the major determinant of the Penner serotyping scheme. This passive slide agglutination typing system was developed in the early 1980's and was recognized for over two decades as the gold standard for C. jejuni typing. A preliminary multiplex PCR technique covering 17 serotypes was previously developed in order to replace this classic serotyping scheme. Here we report the completion of the multiplex PCR technology that is able to identify all the 47 Penner serotypes types known for C. jejuni. The number of capsule types represented within the 47 serotypes is 35. We have applied this method to a collection of 996 clinical isolates from Thailand, Cambodia and Nepal and were able to successfully determine capsule types of 98% of these. This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Figures

  • Fig 1. This cartoon summarizes the Penner types association into complexes commonly found in literature. The variable CPS biosynthesis loci of type strains belonging to the same complex present a high degree of homology [31, 32].
  • Table 1. Bacterial strains used for analysis and validation of the capsule multiplex PCR.
  • Fig 2. Illustration of theC. jejuiCPS loci described in this study. Putative gene functions were assigned via homology to a protein database by BLAST analyses.
  • Table 2. Clinical strains used to test the multiplex PCR technique.
  • Table 3. Summary of the primer sequences included in theC. jejuni capsule multiplex typing scheme.
  • Table 3. (Continued)
  • Fig 3. Illustration of PCR amplicons expected when using the updatedC. jejuniCPSmultiplex PCR. Lane 1, 100-bp NEB DNA standard; lane 2, mixture of PCR products obtained with all the templates from the alpha mix; lane 3, 100-bp NEB DNA standard; lane 4, mixture of PCR products obtained with all the templates from the beta mix; lane 5, 100-bp NEB DNA standard; lane 6, mixture of PCR products with all the templates from the gammamix; lane 7, 100-bp NEB DNA standard; lane 8, mixture of PCR products with all the templates from the gammamix.
  • Table 4. Summary of theC. jejuni capsule multiplex PCR expected results.

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CITATION STYLE

APA

Poly, F., Serichantalergs, O., Kuroiwa, J., Pootong, P., Mason, C., Guerry, P., & Parker, C. T. (2015). Updated campylobacter jejuni capsule PCR multiplex typing system and its application to clinical isolates from south and southeast Asia. PLoS ONE, 10(12). https://doi.org/10.1371/journal.pone.0144349

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