Development of a multiplex taqMan real-time PCR assay for typing of Mycoplasma pneumoniae based on type-specific indels identified through whole genome sequencing

Bernard J Wolff; Alvaro J Benitez; Heta P Desai; Shatavia S Morrison; Maureen H Diaz; Jonas M Winchell

doi:10.1016/j.diagmicrobio.2016.11.013

Development of a multiplex taqMan real-time PCR assay for typing of Mycoplasma pneumoniae based on type-specific indels identified through whole genome sequencing

Diagn Microbiol Infect Dis. 2017 Mar;87(3):203-206. doi: 10.1016/j.diagmicrobio.2016.11.013. Epub 2016 Nov 25.

Authors

Bernard J Wolff¹, Alvaro J Benitez¹, Heta P Desai¹, Shatavia S Morrison¹, Maureen H Diaz¹, Jonas M Winchell²

Affiliations

¹ Division of Bacterial Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
² Division of Bacterial Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA. Electronic address: jwinchell@cdc.gov.

PMID: 27923522
DOI: 10.1016/j.diagmicrobio.2016.11.013

Abstract

We developed a multiplex real-time PCR assay for simultaneously detecting M. pneumoniae and typing into historically-defined P1 types. Typing was achieved based on the presence of short type-specific indels identified through whole genome sequencing. This assay was 100% specific compared to existing methods and may be useful during epidemiologic investigations.

Keywords: M. pneumoniae; Next generation sequencing; P1 typing; Real-time PCR.

Published by Elsevier Inc.

MeSH terms

Base Sequence
DNA, Bacterial / genetics
Genome, Bacterial / genetics*
High-Throughput Nucleotide Sequencing / methods*
Humans
Molecular Typing / methods*
Mycoplasma pneumoniae / classification
Mycoplasma pneumoniae / genetics*
Pneumonia, Mycoplasma / diagnosis*
Pneumonia, Mycoplasma / microbiology
Real-Time Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Sequence Alignment
Sequence Analysis, DNA / methods*

Substances

DNA, Bacterial