Validation of a previously developed quantitative polymerase chain reaction for the detection and quantification of<i>Mycoplasma synoviae</i>in chicken joint specimens

Dijkman, Remco; Feberwee, Anneke; Landman, W. J. M.

doi:10.1080/03079457.2013.766669

Cited by 7 publications

(8 citation statements)

References 44 publications

(46 reference statements)

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“…Until now, MS culturing methods, serological assays and conventional PCRs or real-time PCRs are useful in detection of MS strains (Lauerman et al 1993 ; Hong et al 2004 ; Hess et al 2007 ; Feberwee et al 2008 ; Dijkman et al 2013 ). However, culturing is very laborious, time consuming and requires specific growth medium and incubation (Ratliff et al 2014 ).…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, using PCR and PCR–RFLP method targeted on single-copy domain of the N-terminal vlhA region, it is possible to differentiate field MS isolates from MS-H vaccine strain (Bayatzadeh et al 2014 ). Based on 16S-23S rRNA intergenic spacer region, it was also possible to quantify MS copy number in samples collected from chicken joints using specific qPCR technique (Raviv and Kleven 2009 ; Dijkman et al 2013 ). An attractive alternative to traditional methods of MS detection is LAMP which is simple to set and requires no additional thermal cyclers (Woźniakowski et al 2012 ).…”

Section: Discussionmentioning

confidence: 99%

“…So far, MS diagnosis has been carried out by bacteriological isolation, serological assays, or polymerase chain reaction and its modification (PCR, multiplex PCR or real-time PCR) (Lauerman et al 1993 ; Hong et al 2004 ; Hess et al 2007 ; Feberwee et al 2008 ; Hammond et al 2009 ; Raviv and Kleven 2009 ; Dijkman et al 2013 , Fraga et al 2013 ; Reck et al 2013 ). The sensitivity of direct MS culturing from affected joints is low and often yields negative results, especially in the sub-chronic stage of the disease (Salisch et al 1998 ).…”

Section: Introductionmentioning

confidence: 99%

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Rapid detection of Mycoplasma synoviae by loop-mediated isothermal amplification

et al. 2014

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Mycoplasma synoviae (MS) remains a serious concern in production of poultry and affects world production of chickens and turkeys. Loop-mediated isothermal amplification (LAMP) of DNA has been recently used for the identification of different economically important avian pathogens. The aim of this study was to develop LAMP for simple and inexpensive detection of MS strains in poultry using specifically designed primers targeting hemagglutin A (vlh) gene. The assay was conducted in a water bath for 1 h at 63 °C. The results were visualized after addition of SYBR Green® fluorescent dye. LAMP was specific exclusively for MS without cross-reactivity with other Mycoplasma species. The sensitivity of LAMP was determined as 10−1 CFU/ml and was 1,000 times higher than MS-specific polymerase chain reaction. LAMP assay was conducted on 18 MS field strains to ensure its reliability and usefulness. This is the first report on LAMP development and application for the rapid detection of MS isolated from chickens. This simple method may be applied by diagnostic laboratories without access to expensive equipment.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Rapid detection of Mycoplasma synoviae by loop-mediated isothermal amplification

et al. 2014

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“…Extraction of DNA was performed with the Kapa Express Extract kit as described previously (Dijkman et al, 2013). PCR efficiency was expressed as 10 (−1/slope) .…”

Section: Mycoplasmamentioning

confidence: 99%

Development, validation and field evaluation of a quantitative real-time PCR able to differentiate between field Mycoplasma synoviae and the MS-H-live vaccine strain

2017

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A quantitative PCR (qPCR) able to differentiate between field Mycoplasma synoviae and MS-H vaccine strain was developed, validated and evaluated. It was developed using nucleotide differences in the obg gene. Analytical specificity and sensitivity assessed using DNA from 194 M. synoviae field samples, three different batches of MS-H vaccine and from 43 samples representing four other avian Mycoplasma species proved to be 100%. The detection limit for field M. synoviae and MS-H vaccine strain was 10 and 10 colony-forming units PCR equivalents/g trachea mucus, respectively. The qPCR was able to detect both, field M. synoviae and MS-H vaccine strain in ratios of 1:100 determined both using spiked and field samples. One hundred and twenty samples from M. synoviae-infected non-vaccinated birds, 110 samples from M. synoviae-vaccinated birds from a bird experiment and 224 samples from M. synoviae negative (serology and PCR) birds were used to determine the relative sensitivity and specificity using a previously described M. synoviae PCR as reference. The relative sensitivity and specificity for field M. synoviae were 95.0% and 99.6%, respectively, and 94.6% and 100% for the MS-H-live vaccine, respectively. Field validation and confirmation by multi locus sequence typing revealed that the qPCR correctly distinguished between MS-H and field M. synoviae. Evaluation of the differentiating M. synoviae qPCR in three commercial flocks suggested transmission of MS-H-live vaccine from vaccinated to non-vaccinated flocks at the same farm. Furthermore, it showed evidence for the colonization with field M. synoviae in MS-H-vaccinated flocks.

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“…The above factors are responsible for the severity of the clinical course of infection, epidemic outbreaks in poultry, increasing antibiotic resistance, and economic losses. So far, the diagnosis of poultry infections has been based on clinical study, and bacteria carriage in bird feathers has not been tested [12].…”

Section: Introductionmentioning

confidence: 99%

Analyses of the Polymorphisms in E. coli Strains Associated with Heat-Shock Proteins Hsp 55 Isolated from Bird Feathers

Cybulska¹,

Mahdi-Oraibi²,

Miskiewicz³

et al. 2018

Application of Genetics and Genomics in Poultry Science

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The bird feathers are often colonized by pathogenic microorganisms including mainly bacteria of the E. coli species. There is a grooving evidence that due to colonization of the pathogenic bacteria after slaughter material may lead to different zoonosis diseases that endanger human health. Poultry diseases are a very important issue both economically and epidemiologically in each country. Currently, in practice, EU postmortem monitoring programs are often used to eliminate breeding poultry infected with different pathogenic microorganisms, e.g., E. coli by introducing mandatory bird vaccination. The article describes the combination of genetic and genomic methods that were used in the analysis of species specificity of strains and their genomes, including specific pathogenic bacteria in bird feathers. The aim of the study was (i) to investigate DNA polymorphisms of the obtained bacterial strains isolated from avian feathers (ii) obtaining recombinant Hsp55 protein and defining its role as a potential component of vaccines used in poultry diseases. For the detection and analysis of DNA polymorphisms, we have optimized a new innovative method based on the deficiencies of three molecular techniques, AFLP, PCR-MP, and PCR MP. This new method can be a useful tool used in the genotyping of bacterial E. coli serotypes present on avian feathers after the slaughter process. It also allows to effectively identify a number of early stages of infectious diseases from heterogeneous avian research material. Amplification of polymorphic regions was achieved by using a lower denaturation temperature of the primers and a reduction in the number of cycles in the classical PCR, which simplifies the procedure, preserving the quality and reproducibility of the obtained results. Research of recombinant Hsp55 protein has allowed us to determine the optimal conditions for its production by the classical methods used in proteomic analysis.

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Validation of a previously developed quantitative polymerase chain reaction for the detection and quantification ofMycoplasma synoviaein chicken joint specimens

Cited by 7 publications

References 44 publications

Rapid detection of Mycoplasma synoviae by loop-mediated isothermal amplification

Rapid detection of Mycoplasma synoviae by loop-mediated isothermal amplification

Development, validation and field evaluation of a quantitative real-time PCR able to differentiate between field Mycoplasma synoviae and the MS-H-live vaccine strain

Analyses of the Polymorphisms in E. coli Strains Associated with Heat-Shock Proteins Hsp 55 Isolated from Bird Feathers

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