• Korean Journal of Veterinary Service
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• v.35 no.4
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• pp.289-294
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• 2012

Mycoplasmas are highly fastidious bacteria, difficult to culture and slow growing. Infections with Mycoplasma species can cause a variety of problems in living organisms and in vitro cell cultures. In this study, we investigated the usefulness of a genus-specific consensus PCR analysis method to detect Mycoplasma species. The developed consensus primer pairs MycoF and MycoR were designed specifically to amplify the 16S ribosomal RNA gene (rRNA) of Mycoplasma species by the optimized PCR system. The developed consensus PCR system effectively amplified 215 bp of Mycoplasma genus-specific region of 16S rRNA. In conclusion, we recommend this consensus PCR for monitoring Mycoplasma species in animals, human and cell culture system.

• Korean Journal of Veterinary Service
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• v.38 no.1
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• pp.31-36
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• 2015

Two cats were obtained from a cat kennel. Over the previous 7 days, the cats had shown cough, anorexia, depression and nasal discharge. In this study, the consensus PCR was able to detect successfully Mycoplasma species in nasal swab samples of the cats. To identify feline mycoplasma species from the lung tissue of the cats with pneumonia, Mycoplasma species-specific PCR reactions were conducted. As the results, we could identify M. felis by the positive amplified DNAs. On the other hand, we could not detect any positive reactions with the PCR reaction for M. arginini, M. canis, M. edwardii, M. cynos, M. gateae, M. maculosum, M. molared, M. opalescens, M. spumans and Mycoplasma HRC-689. In conclusion, we detected M. felis from the kenneled cats with pneumonia. We suggested that this consensus PCR would be useful and effective for monitoring Mycoplasma species in various kinds of animals including cats. The application of preceding consensus PCR before the species-specific PCRs may be the most recommended strategy for the identification of Mycoplasma spp.

• Korean Journal of Veterinary Service
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• v.37 no.4
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• pp.247-252
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• 2014

The economic impact of swine mycoplasma infection is high. An accurate diagnosis is often difficult and time consuming. We report the development and validation of an effective multiplex polymerase chain reaction (PCR) assay that detects Mycoplasma (M.) hyopneumoniae and M. hyorhinis. The multi detection of M. hyopneumoniae and M. hyorhinis primer set were employed to detect mycoplasma species and typing of the species was performed on the basis of sequence analysis of the PCR product. The target nucleic acid fragments were specifically amplified by M. hyopneumoniae and M. hyorhinis PCR with 16S ribosomal DNA primers. Single and mixed Mycoplasma species DNA templates were used to evaluate the specificity of the multiplex assay. The corresponding specific DNA products were amplified for each pathogen. The multiplex PCR assay provides a novel tool for simultaneous detection and differentiation of M. hyopneumoniae and M. hyorhinis.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.90-95
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• 1999

A species-specific 760 base pair(bp) BamHI to EcoRI DNA fragment(fMG-2) of lipoprotein gene was isolated from a Mycoplasma gallisepticum(M gallisepticum) genomic library. Based on the DNA sequence data of fMG-2, a pair of 25bp primers was synthesized. When used in the polymerase chain reaction(PCR), 732bp DNA products were amplified from 6 standard strains and 10 field isolates of M gallisepticum, but not from 2 Mycoplasma synoviae and 7 other Mycoplasma species. The lower detection limit was 100fg of the genomic DNA. Identity of the PCR products was confirmed by comparison of patterns of restriction endonuclease analysis with AseI, DraI, EcoRV and SspI.

• Journal of Veterinary Clinics
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• v.33 no.3
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• pp.145-150
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• 2016

We determined the prevalence of feline hemotropic mycoplasma species including 'Candidatus Mycoplasma haemominutum', Mycoplasma haemofelis, and 'Candidatus Mycoplasma turicensis' in naturally infected feral cats in Jeonju, Korea. Forty six feral cats were evaluated by PCR assay targeting the 16S rRNA gene sequence. Nine cats (19.6%) were positive for 'Candidatus Mycoplasma haemominutum', 2 cats (4.3%) were positive for 'Mycoplasm a haemofelis', and 1 cat (2.2%) was infected with both 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemofelis. 'Candidatus Mycoplasma turicensis' was undetected. Partial 16S rRNA gene sequences of Mycoplasma haemofelis were closely (> 96%) related to those from other countries. The amplification of hemoplasma DNA in these samples confirmed the presence of 'Candidatus M. haemominutum' and M. haemofelis in Korea.

• Journal of Veterinary Clinics
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• v.35 no.6
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• pp.273-275
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• 2018

A 6-month-old male cat was presented for investigation of depression, loss of appetite, dehydration, pale conjunctival mucous membrane, weight loss, fast heart and respiratory rates, nasal discharge and cough. Nasal swabs collected from the studied cat. As the results of bacterial culture with nasal swabs, it was suspected with Mycoplasma spp. Also, Mycoplasma species was detected by the PCR reaction with Mycoplasma genus primers. At species PCR assay, the specimens evaluated for the presence of M. felis, M. arginini, M. gateae, and Acholeplasma laidlawii and the result was visualization of bands from 238 bp in agarose gel 1.5% showing M. felis amplicons in samples. In conclusion, we detected M. felis in a cat with respiratory disease. PCR was able to detect successfully M. felis infection in cats.

• Journal of Food Hygiene and Safety
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• v.33 no.4
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• pp.306-309
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• 2018

The present study was conducted to investigate the prevalence of Mycoplasma spp. in cows and pigs with pneumonic lung lesions in Gyeonggi province in 2017. One hundred ninety two and 257 lung tissues were collected from slaughtered cows and pigs with pneumonic lesions, respectively, and examined for the presence of Mycoplasma spp. by a genus specific PCR. Among the examined animals, 147 cows (76.5%) and 203 pigs (80.9%) were found to be infected with Mycoplasma spp.. The infected tissues were further examined to identify the specific species of Mycoplasma using species specific PCRs. The only identified species in cows was M. agalactiae which was detected from 16 cows (8.3%), whereas M. dispar, M. bovis, and M. bovirhinis were not detected. In pigs, M. hyopneumoniae was detected from 74 pigs (28.8%) and M. hyorhinis from 13 pigs (5.1%). M. hyosynoviae was not detected. Taken together, the current study indicates Mycoplasma spp. are commonly associated with lung infection in cows and pigs in Korea. Further studies are needed to evaluate the impact of mycoplasma infection on the development of lung diseases in farm animals.

• KSBB Journal
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• v.29 no.5
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• pp.361-371
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• 2014

Mycoplasma is well recognized as one of the most prevalent and serious microbial contaminants of biologic manufacturing processes. Conventional methods for mycoplasma testing, direct culture method and indirect indicator cell culture method, are lengthy, costly and less sensitive to noncultivable species. In this report, we describe a new TaqMan probe-based real-time PCR method for rapid and quantitative detection of mycoplasma contamination during manufacture of biologics. Universal mycoplasma primers were used for mycoplasma PCR and mycoplasma DNA was quantified by use of a specific TaqMan probe. Specificity, sensitivity, and robustness of the real-time PCR method was validated according to the European Pharmacopoeia. The validation results met required criteria to justify its use as a replacement for the culture method. The established real-time PCR assay was successfully applied to the detection of mycoplasma from human keratinocyte and mesenchymal stem cell as well as Vero cell lines artificially infected with mycoplasma. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of mycoplasma contamination during manufacture of biologics.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.6
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• pp.3425-3428
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• 2013

Approximately, 15-20% of all cancers worldwide are caused by infectious agents. Understanding the role of infectious agents on cancer development might be useful for developing new approaches to its prevention. Mycoplasma genitalium is a clinically important sexually transmitted pathogen that has been associated with several human diseases. There have been a few studies suggestive of probable roles of Mycoplasma genitalium in cancer development, including prostate and ovarian cancers and lymphomas, but the role of this microorganism like other Mycoplasma species in neoplasia is still conjectural. Considering the prevalence of Mycoplasma genitalium infections and also the emergence of resistant strains, Mycoplasma genitalium needs more attention in the infectious agent cancer-causing research area.

• Korean Journal of Veterinary Service
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• v.38 no.2
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• pp.101-106
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• 2015

Murine mycoplasmosis, caused by Mycoplasma (M.) pulmonis, is a prominent disease in rodent animals. The aim of this study was to develop a sensitive and specific PCR assay to detect M. pulmonis in animals and to assess the suitability of this assay for the detection of mycoplasmal infection in rats experimentally infected with M. pulmonis. A new PCR assay using the M. pulmonis-specific primer pairs MPul-F and MPul-R was developed. The primers and probe for the assay were designed from regions in the 16S rRNA gene that are unique to M. pulmonis. The novel PCR assay was very specific and sensitive for M. pulmonis, detecting the equivalent of 5 pg of target template DNA. It detected only M. pulmonis and no other Mycoplasma species or other bacterial species. The newly developed PCR assay also effectively detected M. pulmonis infection in rats. These results suggest that this PCR assay using M. pulmonis-specific primer pairs of MPul-F and MPul-R will be useful and effective for monitoring M. pulmonis infection in animals.