Detection ofMycoplasmain Avian Live Virus Vaccines by Polymerase Chain Reaction

Kojima, Akira; Takahashi, Toshio; Kijima, Mayumi; Ogikubo, Yasuaki; Nishimura, Makoto; Nishimura, Shinzo; Harasawa, Ryô; Tsutsumi, Yutaka

doi:10.1006/biol.1997.0108

Cited by 21 publications

(30 citation statements)

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“…A pair of Mycoplasma genus specific primers was designed for the first step of screening and the discrimination from other probable bacteria. 16SrRNA gene was selected as the target gene for detecting the genus, as described by Kojima (Kojima et al, 1997). Then two pairs of M. bovis species specific primers on P48 and uvrC genes were designed as described by Fu and Sabramaniam, respectively (Fu et al, 2014;Sabramaniam et al, 1998) (Table 1).…”

Section: Dna Extraction and Primers Designmentioning

confidence: 99%

“…No. DP1603) (Cinnagen, Iran), 1.5 mM MgCl 2 in an automated Thermo cycler (FALC, Germany) under the following programs: a) for the 16SrRNA encoding gene (Kojima et al, 1997), 94 °C (7.5min) of initial denaturation and then 30 cycles of denaturation at 94 °C (30s), annealing at 56 °C (30s), elongation at 72 °C (60s) and final extension in 72 °C (5min). b) for the amplification of P48 encoding gene (Fu et al, 2014) Another PCR with species specific designed primers on the uvrC gene, was performed on randomly selected samples which proved as M. bovis by P48 specific primers as mentioned above.…”

Section: Mycoplasmamentioning

confidence: 99%

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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: Dna Extraction and Primers Designmentioning

confidence: 99%

Section: Mycoplasmamentioning

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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“…Despite precautionary measures and systematic mycoplasma monitoring, mycoplasma contamination is periodically detected in veterinary and human live virus vaccines or viral stocks produced by multiple manufacturers worldwide (4,5,10,11,28,30,34,46,55,66). Other biologics (nonvaccine products, e.g., commercial diagnostic antigens) were also reported to be contaminated with mycoplasmas (37,60).…”

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confidence: 99%

Biological Enrichment of Mycoplasma Agents by Cocultivation with Permissive Cell Cultures

Volokhov

Kong

George

et al. 2008

Appl Environ Microbiol

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In this study, we describe our results on the evaluation of the ability of different permissive mammalian cell lines to support the biological enrichment of mycoplasma species known to be bacterial contaminants of cell substrates. The study showed that this approach is able to significantly improve the efficiency of mycoplasma detection based on nucleic acid testing or biochemical technologies (e.g., MycoAlert mycoplasma detection). each mycoplasma species demonstrated that these common cell line contaminants can be detected reliably after 7-day enrichment in MDCK cell culture at contamination levels of 0.05 to 0.25 CFU/ml. The High Five insect cell line was also found to be able to support the efficient growth of most mycoplasma species tested, except for M. hyorhinis strain DBS1050. However, mycoplasma growth in insect cell culture was demonstrated to be temperature dependent, and the most efficient growth was observed when the incubation temperature was increased from 28°C to between 35 and 37°C. We believe that this type of mycoplasma enrichment is one of the most promising approaches for improving the purity and safety testing of cell substrates and other cell-derived biologics and pharmaceuticals.

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“…DNA was extracted from samples by conventional procedures (10) and examined by PCR to identify genus-positive samples. All genus-positive samples were tested by a specific PCR procedure for M. mycoides cluster and Ma.…”

Section: Methodsmentioning

confidence: 99%

Mixed infection zones may be important in the epidemiology of contagious agalactia

Jafarizadeh

Pourbakhsh

Tadayon

et al. 2016

Journal of Veterinary Research

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Introduction: The current study was designed to detect Mycoplasma agalactiae (Ma), Mycoplasma mycoides subsp. capri (Mmc), Mycoplasma capricolum subsp. capricolum (Mcc) and Mycoplasma putrefaciens (Mp) in sheep and goats with clinical signs consistent with contagious agalactia. Material and Methods: A total of 299 samples were collected from 55 mixed herds in Azarbaijan-e-Sharghi province, Iran. Samples were examined using PCR and culture methods. Results: The findings showed that in 40 herds at least one sample was positive by PCR or culture method. Moreover, out of 274 sheep samples, 101 were proved to be positive using the PCR technique and 76 were found positive using the culture method. Out of 25 goat samples, 10 were found positive using PCR and 9 were positive through the culture method. Less than 20% of isolated mycoplasmas were Ma. Ma was detected from almost all studied regions in the province while Mmc, Mcc, and Mp were detected only in a very limited area that was deemed by the research group the mixed infection zone. Conclusion: In vaccination or eradication projects, it would be more economical to focus on mixed infection zones. Further investigation on mixed infection zones could facilitate better understanding of contagious agalactia epidemiology.

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Detection ofMycoplasmain Avian Live Virus Vaccines by Polymerase Chain Reaction

Cited by 21 publications

References 0 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

Biological Enrichment of Mycoplasma Agents by Cocultivation with Permissive Cell Cultures

Mixed infection zones may be important in the epidemiology of contagious agalactia

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