Rapid differential diagnosis of Mycoplasma agalactiae and Mycoplasma bovis based on a multiplex-PCR and a PCR-RFLP

Antonio Foddai; Graziano Idini; Marisa Fusco; Nives Rosa; Cristian De la Fe; Susanna Zinellu; S Corona; Sebastiana Tola

doi:10.1016/j.mcp.2004.11.007

Rapid differential diagnosis of Mycoplasma agalactiae and Mycoplasma bovis based on a multiplex-PCR and a PCR-RFLP

Antonio Foddai, Graziano Idini, Marisa Fusco, Nives Rosa, Cristian De la Fe, Susanna Zinellu, S Corona, Sebastiana Tola

Research output: Contribution to journal › Article › peer-review

Abstract

The membrane-protein 81 gene (mb-mp81) of Mycoplasma bovis was cloned, sequenced and compared to membrane-protein 81 gene (ma-mp81) of Mycoplasma agalactiae. After alignment of both sequences, specific primers pairs were designed from variable or unchanging nucleotide segments. In this study, we describe the development and optimization of a multiplex-PCR (MPCR) for the rapid detection of M. agalactiae and M. bovis strains. In addition, a simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the restriction enzymes AluI, DraI, RsaI and XbaI, is described to distinguish between both species. The results suggest that MPCR and PCR-RFLP assays could be used as an alternative method in routine diagnosis for rapid and specific simultaneous detection of M. agalactiae and M. bovis.

Original language	English
Pages (from-to)	207-212
Journal	Molecular and Cellular Probes
Volume	19
Issue number	3
DOIs	https://doi.org/10.1016/j.mcp.2004.11.007
Publication status	Published - Jun 2005
Externally published	Yes

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@article{47730648b8be4858a43200e6e377d2a5,

title = "Rapid differential diagnosis of Mycoplasma agalactiae and Mycoplasma bovis based on a multiplex-PCR and a PCR-RFLP",

abstract = "The membrane-protein 81 gene (mb-mp81) of Mycoplasma bovis was cloned, sequenced and compared to membrane-protein 81 gene (ma-mp81) of Mycoplasma agalactiae. After alignment of both sequences, specific primers pairs were designed from variable or unchanging nucleotide segments. In this study, we describe the development and optimization of a multiplex-PCR (MPCR) for the rapid detection of M. agalactiae and M. bovis strains. In addition, a simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the restriction enzymes AluI, DraI, RsaI and XbaI, is described to distinguish between both species. The results suggest that MPCR and PCR-RFLP assays could be used as an alternative method in routine diagnosis for rapid and specific simultaneous detection of M. agalactiae and M. bovis.",

author = "Antonio Foddai and Graziano Idini and Marisa Fusco and Nives Rosa and {De la Fe}, Cristian and Susanna Zinellu and S Corona and Sebastiana Tola",

year = "2005",

month = jun,

doi = "10.1016/j.mcp.2004.11.007",

language = "English",

volume = "19",

pages = "207--212",

journal = "Molecular and Cellular Probes",

issn = "0890-8508",

publisher = "Academic Press Inc.",

number = "3",

}

TY - JOUR

T1 - Rapid differential diagnosis of Mycoplasma agalactiae and Mycoplasma bovis based on a multiplex-PCR and a PCR-RFLP

AU - Foddai, Antonio

AU - Idini, Graziano

AU - Fusco, Marisa

AU - Rosa, Nives

AU - De la Fe, Cristian

AU - Zinellu, Susanna

AU - Corona, S

AU - Tola, Sebastiana

PY - 2005/6

Y1 - 2005/6

N2 - The membrane-protein 81 gene (mb-mp81) of Mycoplasma bovis was cloned, sequenced and compared to membrane-protein 81 gene (ma-mp81) of Mycoplasma agalactiae. After alignment of both sequences, specific primers pairs were designed from variable or unchanging nucleotide segments. In this study, we describe the development and optimization of a multiplex-PCR (MPCR) for the rapid detection of M. agalactiae and M. bovis strains. In addition, a simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the restriction enzymes AluI, DraI, RsaI and XbaI, is described to distinguish between both species. The results suggest that MPCR and PCR-RFLP assays could be used as an alternative method in routine diagnosis for rapid and specific simultaneous detection of M. agalactiae and M. bovis.

AB - The membrane-protein 81 gene (mb-mp81) of Mycoplasma bovis was cloned, sequenced and compared to membrane-protein 81 gene (ma-mp81) of Mycoplasma agalactiae. After alignment of both sequences, specific primers pairs were designed from variable or unchanging nucleotide segments. In this study, we describe the development and optimization of a multiplex-PCR (MPCR) for the rapid detection of M. agalactiae and M. bovis strains. In addition, a simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the restriction enzymes AluI, DraI, RsaI and XbaI, is described to distinguish between both species. The results suggest that MPCR and PCR-RFLP assays could be used as an alternative method in routine diagnosis for rapid and specific simultaneous detection of M. agalactiae and M. bovis.

U2 - 10.1016/j.mcp.2004.11.007

DO - 10.1016/j.mcp.2004.11.007

M3 - Article

SN - 0890-8508

VL - 19

SP - 207

EP - 212

JO - Molecular and Cellular Probes

JF - Molecular and Cellular Probes

IS - 3

ER -

Rapid differential diagnosis of Mycoplasma agalactiae and Mycoplasma bovis based on a multiplex-PCR and a PCR-RFLP

Abstract

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