Related Experiment Video
Updated: Jul 11, 2026
Optimized PCR-based Detection of Mycoplasma
Published on: June 20, 2011
An evaluation of PCR methods to detect strains of Mycoplasma fermentans
Baharak Afshar1, David Pitcher, Robin A J Nicholas
1Division of Life Sciences, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 8WA, UK. baharak.afshar@hpa.org.uk
Abstract:
A panel of 30 putative Mycoplasma fermentans strains, isolated from various sources including human, ovine and cell lines, were tested by a previously described polymerase chain reaction (PCR) to confirm their identity by amplification of a conserved 206 bp region of the insertion sequence IS1550. In addition, the application of another PCR based on the major part of the IS1550 element showed one or two products of different length (1144 and 1341 bp) enabling M. fermentans strains to be divided into two types designated as Type A and Type B. A PCR, which amplifies the macrophage activating lipopeptide gene (malp), supported the identification of all the strains as M. fermentans. Thirteen other species of Mycoplasma from human sources gave negative results in these tests, with the exception of Mycoplasma orale, which was detected by both IS1550-PCRs based on the major part and the conserved 206 bp region of the IS1550 element. This study suggests that all M. fermentans isolates possess both the IS1550 element and the malp gene. In contrast to the IS1550, the malp gene is shown to be species-specific and the use of a malp PCR described here could prove to be a useful adjunct to IS1550 detection as confirmation of the presence of M. fermentans in clinical material.
Insights
This study confirms Mycoplasma fermentans identification using polymerase chain reaction (PCR) targeting the IS1550 element and macrophage activating lipopeptide (malp) gene. The malp gene PCR offers species-specific confirmation for Mycoplasma fermentans detection.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Mycoplasma fermentans is a pathogen with diverse isolates.
- Accurate identification methods are crucial for clinical and research settings.
Purpose of the Study:
- To develop and validate PCR-based methods for accurate Mycoplasma fermentans identification.
- To differentiate Mycoplasma fermentans strains and assess species-specificity of diagnostic targets.
Main Methods:
- Polymerase chain reaction (PCR) targeting the conserved 206 bp region of IS1550.
- PCR targeting the major part of IS1550 to differentiate strains into Type A and Type B.
- PCR targeting the macrophage activating lipopeptide (malp) gene for species-specific identification.
Main Results:
- All 30 Mycoplasma fermentans strains were confirmed using IS1550 and malp gene PCR.
- IS1550 PCR differentiated strains into Type A and Type B based on product length.
- Mycoplasma orale was detected by IS1550 PCRs, but malp PCR was specific to Mycoplasma fermentans.
- All M. fermentans isolates possessed both the IS1550 element and the malp gene.
Conclusions:
- The IS1550 element and malp gene are present in all Mycoplasma fermentans isolates.
- The malp gene PCR is a species-specific tool for confirming Mycoplasma fermentans.
- Combining IS1550 and malp gene PCR provides robust identification and differentiation of Mycoplasma fermentans.
Related Concept Videos
02:57Real Time RT-PCR
The real-time quantification of the number of amplified products is...
01:30Methods to Assess Microbial Populations