Polymerase Chain Reaction Optimization for Mycoplasma gallisepticum and M. synoviae Diagnosis

Silveira, Ricardo M.; Fiorentin, L.; Marques, Edmundo Kanan

doi:10.2307/1592392

Cited by 17 publications

(13 citation statements)

References 10 publications

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“…Some papers with MS have been published in Brazil, such as of economical impacts (Balem & Fiorentin 1990), pathogenicity determination of autochthonous MS strains in chicken embryos (Fiorentin & Jeanisch 1994), MS diagnosis establishment by PCR (Silveira et al 1996), and MS and MG eradication from flocks of breeder hens with antibiotic treatment of eggs, in combination with other control measures (Fiorentin & Jeanisch 1994, Nascimento & Nascimento 1994.…”

Section: Discussionmentioning

confidence: 99%

Epidemiological survey on Mycoplasma gallisepticum and M. synoviae by multiplex PCR in commercial poultry

et al. 2009

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ABSTRACT.-Buim M.R., Mettifogo E., Timenetsky J., Kleven S. & Ferreira A.J.P. 2009 Mycoplasmas are important avian pathogens, which cause respiratory and joint diseases that result in large economic losses in Brazilian and world-wide poultry industry. This investigation regarding the main species of mycoplasmas, Mycoplasma gallisepticum (MG) and M. synoviae (MS), responsible for the above mentioned conditions, was carried out through PCR Multiplex analysis. One thousand and forty-six (1,046) samples of tracheal swabs and piped embryos were collected from 33 farms with laying hens, breeders, broilers or hatchery, located in the Brazilian states of São Paulo, Paraná and Pernambuco, where respiratory problems or drops in egg production had occurred. The MG and MS prevalence on the farms was 72.7%. These results indicated (1) high dissemination of mycoplasmas in the evaluated farms, with predominance of MS, either as single infectious agent or associated with other mycoplasmas in 20 farms (60.6%), and (2) an increase of MS and decrease of MG infection in Brazilian commercial poultry. Epidemiological survey on Mycoplasma gallisepticum and M. synoviae by multiplex PCR in commercial poultry 553

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Section: Discussionmentioning

confidence: 99%

Epidemiological survey on Mycoplasma gallisepticum and M. synoviae by multiplex PCR in commercial poultry

et al. 2009

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“…However, the latter is generally expensive and time consuming (requiring 1-2 weeks to complete). Recently, PCR has been employed for identification of suspected cultures or for rapid detection of M. synoviae directly from clinical samples (Garcia et al, 1995;Kiss et al, 1997;Lauerman et al, 1993;Silveira et al, 1996;Wang et al, 1997). Further identification of the M. synoviae isolates (i.e.…”

Section: Introductionmentioning

confidence: 99%

Classification of Mycoplasma synoviae strains using single-strand conformation polymorphism and high-resolution melting-curve analysis of the vlhA gene single-copy region

et al. 2007

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Mycoplasma synoviae is an economically important pathogen of poultry worldwide, causing respiratory infection and synovitis in chickens and turkeys. Identification of M. synoviae isolates is of critical importance, particularly in countries in which poultry flocks are vaccinated with the live attenuated M. synoviae strain MS-H. Using oligonucleotide primers complementary to the single-copy conserved 59 end of the variable lipoprotein and haemagglutinin gene (vlhA), amplicons of~400 bp were generated from 35 different M. synoviae strains/isolates from chickens and subjected to mutation scanning analysis. Analysis of the amplicons by single-strand conformation polymorphism (SSCP) revealed 10 distinct profiles (A-J). Sequencing of the amplicons representing these profiles revealed that each profile related to a unique sequence, some differing from each other by only one base-pair substitution. Comparative high-resolution melting (HRM) curve analysis of the amplicons using SYTO 9 green fluorescent dye also displayed profiles which were concordant with the same 10 SSCP profiles (A-J) and their sequences. For both mutation detection methods, the Australian M. synoviae strains represented one of the A, B, C or D profiles, while the USA strains represented one of the E, F, G, H, I or J profiles. The results presented in this study show that the PCR-based SSCP or HRM curve analyses of vlhA provide high-resolution mutation detection tools for the detection and identification of M. synoviae strains. In particular, the HRM curve analysis is a rapid and effective technique which can be performed in a single test tube in less than 2 h.

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“…Unfortunately, due to the difficulty of isolating and culturing M. gallisepticum from chickens, little is known about the in vivo PK-PD profiles of valnemulin against M. gallisepticum. Molecular methods, such as hybridization techniques, PCR, or combinations of both techniques to detect the presence of bacterium-specific DNA sequences, were used in both qualitative and quantitative detection of the organism (8,9). Recently, real-time PCR (RT-PCR) methods that monitor the production of specific PCR products by fluorimetric detection of amplified products and use melting curve analysis or specific hybridization probes for identification have been used for qualitative and quantitative detection of various pathogens (10,11).…”

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confidence: 99%

In Vivo Pharmacokinetic/Pharmacodynamic Profiles of Valnemulin in an Experimental Intratracheal Mycoplasma gallisepticum Infection Model

Xiao

Sun

Fang

et al. 2015

Antimicrob Agents Chemother

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The AUC 24 /MIC ratios for mycoplasmastasis (a reduction of 0 log 10 color-changing unit [CCU] equivalents/ml), a reduction of 1 log 10 CCU equivalents/ml, and a reduction of 2.5 log 10 CCU equivalents/ml are 28,820, 38,030, and 56,256, respectively. In addition, we demonstrated that valnemulin at a dose of 6.5 mg/kg resulted in a reduction of 2.5 log 10 CCU equivalents/ml. These investigations provide a solid foundation for the usage of valnemulin in poultry with M. gallisepticum infections.

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Polymerase Chain Reaction Optimization for Mycoplasma gallisepticum and M. synoviae Diagnosis

Cited by 17 publications

References 10 publications

Epidemiological survey on Mycoplasma gallisepticum and M. synoviae by multiplex PCR in commercial poultry

Epidemiological survey on Mycoplasma gallisepticum and M. synoviae by multiplex PCR in commercial poultry

Classification of Mycoplasma synoviae strains using single-strand conformation polymorphism and high-resolution melting-curve analysis of the vlhA gene single-copy region

In Vivo Pharmacokinetic/Pharmacodynamic Profiles of Valnemulin in an Experimental Intratracheal Mycoplasma gallisepticum Infection Model

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