Specific real-time PCR assays for the detection and quantification of Mycoplasma mycoides subsp. mycoides SC and Mycoplasma capricolum subsp. capripneumoniae

Lorenzon, Sophie; Manso-Silván, Lucía; Thiaucourt, François

doi:10.1016/j.mcp.2008.07.003

Cited by 35 publications

(33 citation statements)

References 17 publications

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“…Moreover, no preculturing or extensive sample preparation is needed. In addition, many sensitive and specific nucleic acid-based detection methods are available for CBPP (32)(33)(34), yet the rather elaborate requirements for the laboratory facilities, sample costs, and processing associated with these techniques have limited their routine use in Africa. Recently, a sensitive and easy-to-use loop-mediated isothermal amplification (LAMP)-based assay for CBPP was published, which overcomes many of the above-mentioned constraints (35).…”

Section: Discussionmentioning

confidence: 99%

Development of a Novel Cocktail Enzyme-Linked Immunosorbent Assay and a Field-Applicable Lateral-Flow Rapid Test for Diagnosis of Contagious Bovine Pleuropneumonia

et al. 2016

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e Contagious bovine pleuropneumonia (CBPP) is a severe respiratory disease that is widespread in sub-Saharan Africa. It is caused by Mycoplasma mycoides subsp. mycoides, a bacterium belonging to the Mycoplasma mycoides cluster. In the absence of an efficient CBPP vaccine, improved and easy-to-use diagnostic assays for recurrent testing combined with isolation and treatment of positive animals represent an option for CBPP control in Africa. Here we describe the comprehensive screening of 17 immunogenic Mycoplasma mycoides subsp. mycoides proteins using well-characterized bovine sera for the development of a novel cocktail enzyme-linked immunosorbent assay (ELISA) for laboratory use. Two recombinant Mycoplasma immunogens, MSC_0136 and MSC_0636, were used to set up a standardized cocktail ELISA protocol. According to the results from more than 100 serum samples tested, the sensitivity and specificity of the novel cocktail ELISA were 85.6% and 96.4%, respectively, with an overall diagnostic accuracy comparable to that of the Office International des Epizooties (OIE)-prescribed serological assays. In addition, we provide a proof of principle for a field-applicable, easy-to-use commercially produced prototype lateral-flow test for rapid (<30-min) diagnosis of CBPP.

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Section: Discussionmentioning

confidence: 99%

Development of a Novel Cocktail Enzyme-Linked Immunosorbent Assay and a Field-Applicable Lateral-Flow Rapid Test for Diagnosis of Contagious Bovine Pleuropneumonia

et al. 2016

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“…Specific detection methods were then developed based on PCR technology and the empirical search for specific restriction sites in the amplified fragment or with PCR primers allowing a specific amplification [8,9]. Finally, MmmSC specific monoclonal antibodies were obtained and, more recently, specific real-time PCR methods have also been validated [10-13]. …”

Section: Introductionmentioning

confidence: 99%

Mycoplasma mycoides, from "mycoides Small Colony" to "capri". A microevolutionary perspective

et al. 2011

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BackgroundThe Mycoplasma mycoides cluster consists of five species or subspecies that are ruminant pathogens. One subspecies, Mycoplasma mycoides subspecies mycoides Small Colony (MmmSC), is the causative agent of contagious bovine pleuropneumonia. Its very close relative, Mycoplasma mycoides subsp. capri (Mmc), is a more ubiquitous pathogen in small ruminants causing mastitis, arthritis, keratitis, pneumonia and septicaemia and is also found as saprophyte in the ear canal. To understand the genetics underlying these phenotypic differences, we compared the MmmSC PG1 type strain genome, which was already available, with the genome of an Mmc field strain (95010) that was sequenced in this study. We also compared the 95010 genome with the recently published genome of another Mmc strain (GM12) to evaluate Mmc strain diversity.ResultsThe MmmSC PG1 genome is 1,212 kbp and that of Mmc 95010 is ca. 58 kbp shorter. Most of the sequences present in PG1 but not 95010 are highly repeated Insertion Sequences (three types of IS) and large duplicated DNA fragments. The 95010 genome contains five types of IS, present in fewer copies than in PG1, and two copies of an integrative conjugative element. These mobile genetic elements have played a key role in genome plasticity, leading to inversions of large DNA fragments. Comparison of the two genomes suggested a marked decay of the PG1 genome that seems to be correlated with a greater number of IS. The repertoire of gene families encoding surface proteins is smaller in PG1. Several genes involved in polysaccharide metabolism and protein degradation are also absent from, or degraded in, PG1.ConclusionsThe genome of MmmSC PG1 is larger than that of Mmc 95010, its very close relative, but has less coding capacity. This is the result of large genetic rearrangements due to mobile elements that have also led to marked gene decay. This is consistent with a non-adaptative genomic complexity theory, allowing duplications or pseudogenes to be maintained in the absence of adaptive selection that would lead to purifying selection and genome streamlining over longer evolutionary times. These findings also suggest that MmmSC only recently adapted to its bovine host.

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“…At CIRAD, DNA was extracted using the DNeasy blood and tissue kit (Qiagen) and tested by real-time PCR for the detection of Mycoplasma capricolum subsp. capripneumoniae (Mccp) (Lorenzon et al, 2008) providing a positive result. Subsequently, a pure mycoplasma culture was isolated from the same sample after four days of incubation in a modified CCPP medium (World Organization for Animal Health, 2009).…”

Section: Resultsmentioning

confidence: 99%

Fatal transmission of contagious caprine pleuropneumonia to an Arabian oryx (Oryx leucoryx)

Chaber¹,

Lignereux²,

Qassimi³

et al. 2014

Veterinary Microbiology

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Specific real-time PCR assays for the detection and quantification of Mycoplasma mycoides subsp. mycoides SC and Mycoplasma capricolum subsp. capripneumoniae

Cited by 35 publications

References 17 publications

Development of a Novel Cocktail Enzyme-Linked Immunosorbent Assay and a Field-Applicable Lateral-Flow Rapid Test for Diagnosis of Contagious Bovine Pleuropneumonia

Development of a Novel Cocktail Enzyme-Linked Immunosorbent Assay and a Field-Applicable Lateral-Flow Rapid Test for Diagnosis of Contagious Bovine Pleuropneumonia

Mycoplasma mycoides, from "mycoides Small Colony" to "capri". A microevolutionary perspective

Fatal transmission of contagious caprine pleuropneumonia to an Arabian oryx (Oryx leucoryx)

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