Conservation of the uvrC gene sequence in Mycoplasma bovis and its use in routine PCR diagnosis

Thomas, Anne; Dizier, Isabelle; Lindén, Annick; Mainil, Jacques; Frey, Joachim; Vilei, Edy M.

doi:10.1016/j.tvjl.2003.10.006

Cited by 14 publications

(14 citation statements)

References 9 publications

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“…Robino also studied the conserved surface lipoprotein P48 as a marker for the detection of M. bovis strains (Robino et al, 2005). One of the housekeeping genes which are known as conserved sequences in organisms, is uvrC encoding gene considered as M. bovis marker's too (Thomas et al, 2004). The uvrC encodes deoxyribodipyrimidine photolyase which removes damaged DNA and play the role in the repairing system in Mycoplasma species (Hotzel et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Isolation, identification and molecular characterization of Mycoplasma bovis in mastitic dairy cattle by PCR and culture methods

Imandar

Pourbakhsh

Jamshidian

et al. 2018

J Hellenic Vet Med Soc

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Mycoplasma bovis is well known as one of the major causative agents of mastitis in dairy cattle herds. The aim of this study was the identification of Mycoplasma bovis strains by PCR and traditional culture methods from a total number of 328 milk samples collected from cows with clinical mastitis symptoms from all over Iran. First step cultures in PPLO broth and agar showed 58 samples (17.69%) as positive. Out of 328 samples, 97 samples (29.57%) were positive for Mycoplasma genus according to the amplification of the 16SrRNA gene performed by PCR and from them, 31 (31.97%) samples were positive by PCR on the P48 gene. The purified P48 positive PCR products were sequenced and results were compared to M. bovis reference strain PG45 (CP002188). A phylogenic tree was created using Neighbor-joining method in MEGA6 software. The studied strain IB220 showed 100% identity with the reference strain of M. bovis and followed the same phylogenetic roots while studied strain IB216 showed 99.7% homology with the reference strain. Twelve selected geographical isolated strains were subjected to Gene Bank under accession numbers of KX772789 to KX772800. This is the first study of the molecular characterization of Mycoplasma bovis in dairy cattle with clinical mastitis from Iran.

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Section: Introductionmentioning

confidence: 99%

Isolation, identification and molecular characterization of Mycoplasma bovis in mastitic dairy cattle by PCR and culture methods

Imandar

Pourbakhsh

Jamshidian

et al. 2018

J Hellenic Vet Med Soc

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“…18 Clinical signs include mastitis, vaginitis, and abortions in adult cattle as well as pneumonia, septic polyarthritis, otitis media, and conjunctivitis in calves. 5,16,20,22 Prevalence of M. bovis is often underestimated in both beef and dairy herds because of the difficulties in diagnosis and the variety of clinical signs associated with this infection and disease caused by this pathogen. 15 Bacterial isolation, considered the gold standard test for mycoplasmal infections, is difficult and time consuming because of the fastidious nature of mycoplasmas, which require 1) specialized media and incubation conditions; 2) extended culture time as a result of the slow growth of the organism (up to 14 days for completion); and 3) additional testing for speciation.…”

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confidence: 99%

Mycoplasma Bovis Real-Time Polymerase Chain Reaction Assay Validation and Diagnostic Performance

et al. 2010

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Abstract. Mycoplasma bovis is an important bacterial pathogen in cattle, producing a variety of clinical diseases. The organism, which requires specialized culture conditions and extended incubation times to isolate and identify, is frequently associated with concurrent infection with other pathogens which can potentially be more easily identified. Real-time polymerase chain reaction (real-time PCR) is a valuable diagnostic technique that can rapidly identify infectious agents in clinical specimens. A real-time PCR assay was designed based on the uvrC gene to identify M. bovis in diagnostic samples. Using culture as the gold standard test, the assay performed well in a variety of diagnostic matrices. 100%]). Low numbers of other sample matrices showed good agreement between results of culture and PCR. A review of clinical cases from 2009 revealed that, in general, PCR was used much more frequently than culture and provided useful diagnostic information in conjunction with clinical signs, signalment, and gross and histopathologic lesions. Diagnostic performance of the real-time PCR assay developed as a testing method indicates that it is a rapid, accurate assay that is adaptable to a variety of PCR platforms and can provide reliable results on an array of clinical samples.

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“…No cross-reactivity was observed with other DNA targets, suggesting 100% specificity of the Taqman assay. The uvrC gene in M. bovis has been shown to be stable and resistant to normal mutation process reinforcing its efficacy as a target for PCR assays [ 16 ]. Thomas et al [ 16 ] have demonstrated the uvrC gene to be a conserved target suitable for PCR based diagnostics by PCR amplification and sequencing from 92 and 20 M. bovis strains respectively.…”

Section: Resultsmentioning

confidence: 99%

“…Subramaniam and colleagues have demonstrated that although the uvrC gene is well conserved in M. bovis and in another small ruminant pathogen Mycoplasma agalactiae , this gene differs significantly between the two phylogenetically closely related Mycoplasma species [ 10 ]. Thomas and colleagues [ 16 ] have demonstrated the uvrC gene to be a conserved target suitable for PCR based diagnostics by PCR amplification and sequencing from ninety-two and twenty M. bovis strains respectively. Interestingly, Marenda and colleagues [ 17 ] have demonstrated the limitation of uvrC target’s universality for M. bovis detection since the uvrC gene sequences are only slightly divergent making it difficult to differentiate M. bovis and M. agalactiae .…”

Section: Introductionmentioning

confidence: 99%

Development and Evaluation of a Novel Taqman Real-Time PCR Assay for Rapid Detection of Mycoplasma bovis: Comparison of Assay Performance with a Conventional PCR Assay and Another Taqman Real-Time PCR Assay

Naikare¹,

Bruno²,

Mahapatra³

et al. 2015

Veterinary Sciences

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The objective of this study was to develop and validate a Taqman real-time PCR assay for the detection of Mycoplasma bovis (M. bovis). Unique primers targeting the highly conserved house-keeping gene (uvrC) were designed and the probe sequence was derived from a previously published microarray study. There was 100% agreement in the outcome between our assay and the other two published assays for M. bovis detection. The analytical limit of detection of our assay is 83 copies of the uvrC gene. This assay was validated on a total of 214 bovine clinical specimens that were submitted to the Texas A&M Veterinary Medical Diagnostic Laboratory (TVMDL), Texas, USA. The specificity of the assay was assessed to be 100% since no cross-reactivity occurred with 22 other bacterial and other Mycoplasma species. We conclude that the uvrC gene serves as a good and reliable diagnostic marker for the accurate and rapid detection of M. bovis from a wider variety of specimen matrices.

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Conservation of the uvrC gene sequence in Mycoplasma bovis and its use in routine PCR diagnosis

Cited by 14 publications

References 9 publications

Isolation, identification and molecular characterization of Mycoplasma bovis in mastitic dairy cattle by PCR and culture methods

Isolation, identification and molecular characterization of Mycoplasma bovis in mastitic dairy cattle by PCR and culture methods

Mycoplasma Bovis Real-Time Polymerase Chain Reaction Assay Validation and Diagnostic Performance

Development and Evaluation of a Novel Taqman Real-Time PCR Assay for Rapid Detection of Mycoplasma bovis: Comparison of Assay Performance with a Conventional PCR Assay and Another Taqman Real-Time PCR Assay

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