Multiplex and quantitative pathogen detection with high-resolution capillary electrophoresis-based single-strand conformation polymorphism

Hee Sung Hwang, Gi Won Shin, Boram Chung, Jeongkyeong Na, Gyoo Yeol Jung

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

8 Scopus citations

Abstract

Among the molecular diagnostic methods for bacteria-induced diseases, capillary electrophoresis-based single-strand conformation polymorphism (CE-SSCP) combined with 16S rRNA gene-specific PCR has enormous potential because it can separate sequence variants using a simple procedure. However, conventional CE-SSCP systems have limited resolution and cannot separate most 16S rRNA gene-specific markers into separate peaks. A high-resolution CE-SSCP system that uses a poly(ethyleneoxide)-poly(propyleneoxide)- poly(ethyleneoxide) triblock copolymer matrix was recently developed and shown to effectively separate highly similar PCR products. In this report, a protocol for the detection of 12 pathogenic bacteria is provided. Pathogen markers were amplified by PCR using universal primers and separated by CE-SSCP; each marker peak was well separated at baseline and showed a characteristic mobility, allowing the easy identification of the pathogens.

Original languageEnglish
Title of host publicationClinical Applications of Capillary Electrophoresis
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages155-163
Number of pages9
ISBN (Print)9781627030281
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume919
ISSN (Print)1064-3745

Keywords

  • 16S rRNA gene
  • CE-SSCP
  • High-resolution
  • Pathogen detection
  • Polymer matrix

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