Molecular typing of Mycoplasma pneumoniae from clinical samples v 1.0
Research type
Research Study
Full title
Molecular typing of Mycoplasma pneumoniae from clinical samples using a Nested modification of the published Multi-Locus Sequence Type (MLST) protocol v 1.0
IRAS ID
214161
Contact name
Vicki Chalker
Contact email
Sponsor organisation
Public Health England
Duration of Study in the UK
0 years, 5 months, 0 days
Research summary
Mycoplasma pneumoniae is an important bacterial pathogen causing respiratory disease in humans of any age that can lead to life-threatening conditions such as extra-pulmonary sequelae (Brown et al., 2015a). Typing of M. pneumoniae is becoming increasingly important due to the global rise in infections and the isolation of macrolide resistant strains. Brown et al. (2015b) developed a multi-locus sequence typing (MLST) scheme for clinical isolates of M. pneumoniae based on the sequences of eight housekeeping genes (ppa, pgm, gyrB, gmk, glyA, atpA, arcC, and adk). However, isolating M. pneumoniae from clinical samples is a laborious specialist process and the need for an adaptation of the developed MLST scheme, for use directly on primary samples, was identified (Brown et al., 2015b).
The aim of this study will be to develop an assay that will incorporate the original MLST primers with a second nested PCR step, designed to increase sensitivity and obtain typing data from lower concentrations of target DNA. The sensitivity of the nested-modified assay will be compared to the original MLST assay, on known concentrations of a M. pneumoniae DNA standard. Once the protocol is optimised, the nested MLST will be tested on DNA extracted from clinical samples and results compared to those obtained from MLST on isolates (where available).REC name
South Central - Oxford B Research Ethics Committee
REC reference
16/SC/0527
Date of REC Opinion
28 Sep 2016
REC opinion
Favourable Opinion