Rapid and accurate typing of Bordetella pertussis targeting genes encoding acellular vaccine antigens using real time PCR and High Resolution Melt analysis

Wai-Fong Chan; Ram P Maharjan; Peter R Reeves; Vitali Sintchenko; Gwendolyn L Gilbert; Ruiting Lan

doi:10.1016/j.mimet.2009.03.007

Rapid and accurate typing of Bordetella pertussis targeting genes encoding acellular vaccine antigens using real time PCR and High Resolution Melt analysis

J Microbiol Methods. 2009 Jun;77(3):326-9. doi: 10.1016/j.mimet.2009.03.007. Epub 2009 Mar 31.

Authors

Wai-Fong Chan¹, Ram P Maharjan, Peter R Reeves, Vitali Sintchenko, Gwendolyn L Gilbert, Ruiting Lan

Affiliation

¹ School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia.

PMID: 19341769
DOI: 10.1016/j.mimet.2009.03.007

Abstract

Real Time-PCR (RT-PCR) and high resolution melt (HRM) analyses were used for rapid typing of genes encoding components of the pertussis acellular vaccine, namely prn, ptxA, fhaB, fim2 and fim3. The length polymorphisms in prn were detected by RT-PCR followed by HRM; single nucleotide polymorphisms in prn and other genes were detected by hairpin primer RT-PCR. These rapid methods are suitable for large-scale studies of vaccine-driven evolution of Bordetella pertussis.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Antigens, Bacterial / genetics*
Antigens, Bacterial / immunology
Bacterial Typing Techniques / methods*
Bordetella pertussis / classification
Bordetella pertussis / genetics*
Bordetella pertussis / immunology
Humans
Pertussis Vaccine / genetics*
Pertussis Vaccine / immunology
Polymerase Chain Reaction / methods*
Polymorphism, Single Nucleotide
Transition Temperature
Vaccines, Acellular / genetics
Vaccines, Acellular / immunology
Whooping Cough / microbiology*

Substances

Antigens, Bacterial
Pertussis Vaccine
Vaccines, Acellular