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Updated: Aug 19, 2026
Optimized PCR-based Detection of Mycoplasma
Published on: June 20, 2011
Inactivation, storage, and PCR detection of Mycoplasma on FTA filter paper
Hugo Moscoso1, Stephan G Thayer, Charles L Hofacre
1Poultry Diagnostic and Research Center, Department of Avian Medicine, University of Georgia, Athens, GA 30602-4875, USA.
Abstract:
We evaluated the feasibility of using Flinders Technology Associates (FTA) filter paper for the inactivation and storage of mycoplasma DNA templates and their detection by the polymerase chain reaction (PCR). FTA paper is a cotton-based cellulose membrane containing lyophilized chemicals that lyses most types of bacteria and viruses. Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) cultures were spotted at various volumes on the filter paper and stored at different temperatures for various periods of time before performing PCR. MG and MS were readily detected at all time frames (1-60 days) independent of the volume applied (1-100 microl) or storage temperature (4 C-41 C). Sensitivity and specificity of the FTA-PCR were comparable to the standard diagnostic PCR, allowing the detection of MG/MS in field samples without interference by nontargeted mycoplasma. Analysis of 193 field samples by both methods showed nearly 100% agreement with serology and culture results. The long-term DNA stability at a wide range of temperatures makes the FTA cards a good alternative for collecting and simultaneously inactivating mycoplasma. It also offers the convenience of storage and transport of DNA in a cost-effective manner for further molecular analysis, such as restriction enzyme length polymorphism and nucleotide sequencing.
Insights
Flinders Technology Associates (FTA) filter paper effectively inactivates and stores mycoplasma DNA for polymerase chain reaction (PCR) detection. This method offers a stable, cost-effective alternative for sample collection and transport in avian diagnostics.
Area of Science:
- Veterinary Microbiology
- Molecular Diagnostics
- Biotechnology
Background:
- Mycoplasma species, such as Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS), are significant avian pathogens.
- Accurate and stable DNA preservation is crucial for reliable molecular diagnostics like polymerase chain reaction (PCR).
- Current methods for sample collection and storage can be cumbersome and may affect DNA integrity.
Purpose of the Study:
- To assess the feasibility of using Flinders Technology Associates (FTA) filter paper for inactivating and storing mycoplasma DNA templates.
- To evaluate the efficacy of FTA paper-based PCR (FTA-PCR) for detecting Mycoplasma gallisepticum and Mycoplasma synoviae.
- To compare the performance of FTA-PCR with standard diagnostic PCR methods.
Main Methods:
- Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) cultures were applied to FTA filter paper in varying volumes.
- Samples on FTA paper were stored at different temperatures (4°C to 41°C) for extended periods (1-60 days).
- DNA was extracted from FTA paper and subjected to PCR for MG/MS detection; results were compared to standard PCR, serology, and culture.
Main Results:
- MG and MS were reliably detected by FTA-PCR across all tested time frames, volumes, and storage temperatures.
- FTA-PCR demonstrated sensitivity and specificity comparable to standard PCR, with no interference from non-targeted mycoplasma.
- Analysis of 193 field samples showed nearly 100% agreement between FTA-PCR, serology, and culture methods.
Conclusions:
- FTA filter paper is a suitable medium for the collection, simultaneous inactivation, and long-term storage of mycoplasma DNA.
- FTA-PCR provides a stable, cost-effective, and convenient method for detecting MG and MS in field samples.
- This approach facilitates further molecular analyses, including restriction enzyme length polymorphism and nucleotide sequencing.
