Real-time polymerase chain reaction assays for the detection of memebers of the Mycoplasma mycoides cluster

Fitzmaurice J., Sewell M.M.H., Manso-Silvan Lucia, Thiaucourt François, McDonald W.L., O'Keefe J.S.. 2008. Real-time polymerase chain reaction assays for the detection of memebers of the Mycoplasma mycoides cluster. New Zealand Veterinary Journal, 56 (1) : 40-47.

Résumé : AIM: To develop real-time PCR assays for the detection and differentiation of members of the Mycoplasma mycoides cluster. METHODS: Five real-time PCR assays were designed to al-low differentiation of members of the M mycoides cluster: an assay for detection of the M mycoides subspecies, viz M mycoides subsp mycoides large colony (MmmLC), M mycoides subsp capri (Mmc), and M mycoides subsp mycoides small colony (MmmSC); one for the detection of the M capricolum subspecies, viz M capricolum subsp capricolum (Mcc), M capricolum subsp capripneumoniae (Mccp), and Mycoplasma sp bovine group 7 (BG7); and three for the specific detection of MmmSC, Mccp, and BG7. A panel of 74 Mycoplasma isolates from various geographical origins and a panel of 21 other bacterial isolates were used to evaluate the sensitivity and specificity of the as-says. RESULTS: The assays displayed 100% analytical sensitivity in detecting all target Mycoplasma isolates. The analytical detection limit for the assays to detect the M mycoides subspecies, M capricolum subspecies, and MmmSC was determined to be 100 fg of genomic DNA, while the Mccp and BG7 assays had a detection limit of 100 fg and 10 fg of genomic DNA, respectively. The M mycoides subspecies assay had a detection limit of 103 (SD 102) cfu/ml milk, 104 (SD 104) cfu per swab, and 103 (SD 103) cfu/g lung in inoculated samples. The assays displayed 100% specificity when applied to non-target bacterial isolates and to 110 culture-negative milk samples. CONCLUSIONS: The assays were highly sensitive and specific, and provide accurate detection and differentiation of the members of the M mycoides cluster.

Mots-clés Agrovoc : Mycoplasma mycoides, identification, PCR, diagnostic, Mycoplasma capricolum

Mots-clés complémentaires : Mycoplasma capripneumoniae

Classification Agris : L72 - Organismes nuisibles des animaux
L50 - Physiologie et biochimie animales

Champ stratégique Cirad : Axe 4 (2005-2013) - Santé animale et maladies émergentes

Auteurs et affiliations

• Fitzmaurice J., Ministry of Agriculture and Forestry (Nouvelle-Zélande) (NZL)
• Sewell M.M.H., Ministry of Agriculture and Forestry (Nouvelle-Zélande) (NZL)
• Manso-Silvan Lucia, CIRAD-BIOS-UPR Contrôle des maladies (FRA) ORCID: 0000-0002-5539-3804
• Thiaucourt François, CIRAD-BIOS-UPR Contrôle des maladies (FRA)
• McDonald W.L., Ministry of Agriculture and Forestry (Nouvelle-Zélande) (NZL)
• O'Keefe J.S., Ministry of Agriculture and Forestry (Nouvelle-Zélande) (NZL)

Source : Cirad - Agritrop (https://agritrop.cirad.fr/543530/)

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