Development and evaluation of a real-time polymerase chain reaction method for the detection of Mycoplasma felis.
Abstract: Infection by Mycoplasma felis is associated with ocular and respiratory disease in cats and respiratory disease in horses. A correct diagnosis is beneficial since the use of specific antimycoplasmal treatment can lead to resolution. The objective of the present study was to develop a real-time polymerase chain reaction (PCR) method based on dual-labeled fluorogenic probe technology, targeting the gene encoding elongation factor Tu (tuf ), for the fast and specific detection of M. felis. Specificity was achieved by basing the assay design on partial sequencing of the tuf gene in strains and clinical isolates of M. felis as well as other mycoplasma species. The detection limit of the developed assay was in the order of 10 copies of target DNA, and no cross-reaction was observed with a panel of several mycoplasma species. Compared to a previously published conventional PCR protocol, the novel assay had equal or slightly improved performance in terms of sensitivity and specificity when analyzing 100 conjunctival swab samples from cats with clinical signs of infection.
Publication Date: 2011-06-15 PubMed ID: 21908343DOI: 10.1177/1040638711407479Google Scholar: Lookup
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- Journal Article
- Clinical Study
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Disease Treatment
- DNA
- Equine Diseases
- Equine Health
- Genetics
- Horses
- Infection
- Infectious Disease
- Laboratory Methods
- Molecular biology
- Mycoplasma
- Polymerase Chain Reaction
- Real-Time PCR
- Respiratory Disease
- Veterinary Care
- Veterinary Medicine
- Veterinary Research
Summary
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The research article is about the development and evaluation of a method that uses real-time polymerase chain reaction (PCR) for detecting the presence of a bacteria called Mycoplasma felis, which is associated with certain diseases in cats and horses.
Research Objectives and Rationale
- The primary objective of this study was to create a real-time PCR method that facilitates rapid and specific detection of the bacterium Mycoplasma felis.
- Accurate diagnosis of the M. felis infection is crucial in providing targeted treatment, which contributes to the resolution of the disease.
- This PCR method was developed using dual-labeled fluorogenic probe technology, targeting a gene in the bacteria known as elongation factor Tu (tuf).
- The significance of this research lies in the fact that M. felis infection can lead to ocular and respiratory conditions in cats and respiratory disease in horses. An efficient and accurate detection method is therefore essential for early diagnosis and treatment.
Methodology
- The specificity of the assay was achieved by designing it based on the partial sequencing of the tuf gene in different strains and clinical isolates of M. felis and other mycoplasma species.
- The detection limit of the developed assay was approximately 10 copies of target DNA, indicating a high level of sensitivity for detecting the bacteria.
- No cross-reaction was observed with a number of other mycoplasma species, further emphasizing the specificity of the assay.
Comparison with Previous Methods
- Results of the newly created real-time PCR method were compared to those of a previously published conventional PCR protocol for performance assessment.
- The novel assay displayed equal or slightly better performance in terms of sensitivity and specificity when evaluated using 100 conjunctival swab samples from cats demonstrating signs of infection.
- This comparison underscores the accuracy and reliability of the new PCR method for detecting M. felis in cats.
Cite This Article
APA
Söderlund R, Bölske G, Holst BS, Aspán A.
(2011).
Development and evaluation of a real-time polymerase chain reaction method for the detection of Mycoplasma felis.
J Vet Diagn Invest, 23(5), 890-893.
https://doi.org/10.1177/1040638711407479 Publication
Researcher Affiliations
- Department of Bacteriology, National Veterinary Institute (SVA), Ulls väg 2B, 75189 Uppsala, Sweden. robert.soderlund@sva.se
MeSH Terms
- Animals
- Cat Diseases / diagnosis
- Cat Diseases / microbiology
- Cats
- Horse Diseases / diagnosis
- Horse Diseases / microbiology
- Horses
- Mycoplasma / classification
- Mycoplasma / isolation & purification
- Mycoplasma Infections / diagnosis
- Mycoplasma Infections / microbiology
- Mycoplasma Infections / veterinary
- Real-Time Polymerase Chain Reaction / methods
- Real-Time Polymerase Chain Reaction / veterinary
- Sensitivity and Specificity
Citations
This article has been cited 5 times.- Ryser-Degiorgis MP, Marti I, Pisano SRR, Pewsner M, Wehrle M, Breitenmoser-Würsten C, Origgi FC, Kübber-Heiss A, Knauer F, Posautz A, Eberspächer-Schweda M, Huder JB, Böni J, Kubacki J, Bachofen C, Riond B, Hofmann-Lehmann R, Meli ML. Management of Suspected Cases of Feline Immunodeficiency Virus Infection in Eurasian Lynx (Lynx lynx) During an International Translocation Program.. Front Vet Sci 2021;8:730874.
- McAloose D, Laverack M, Wang L, Killian ML, Caserta LC, Yuan F, Mitchell PK, Queen K, Mauldin MR, Cronk BD, Bartlett SL, Sykes JM, Zec S, Stokol T, Ingerman K, Delaney MA, Fredrickson R, Ivančić M, Jenkins-Moore M, Mozingo K, Franzen K, Bergeson NH, Goodman L, Wang H, Fang Y, Olmstead C, McCann C, Thomas P, Goodrich E, Elvinger F, Smith DC, Tong S, Slavinski S, Calle PP, Terio K, Torchetti MK, Diel DG. From People to Panthera: Natural SARS-CoV-2 Infection in Tigers and Lions at the Bronx Zoo.. mBio 2020 Oct 13;11(5).
- Tallmadge RL, Anderson R, Mitchell PK, Forbes ZC, Werner B, Gioia G, Moroni P, Glaser A, Thachil AJ, Goodman LB. Characterization of a novel Mycoplasma cynos real-time PCR assay.. J Vet Diagn Invest 2020 Nov;32(6):793-801.
- Spiri AM, Meli ML, Riond B, Herbert I, Hosie MJ, Hofmann-Lehmann R. Environmental Contamination and Hygienic Measures After Feline Calicivirus Field Strain Infections of Cats in a Research Facility.. Viruses 2019 Oct 17;11(10).
- Berger A, Willi B, Meli ML, Boretti FS, Hartnack S, Dreyfus A, Lutz H, Hofmann-Lehmann R. Feline calicivirus and other respiratory pathogens in cats with Feline calicivirus-related symptoms and in clinically healthy cats in Switzerland.. BMC Vet Res 2015 Nov 13;11:282.
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