A specific PCR for the identification of Mycoplasma capricolum subsp. capripneumoniae, the causative agent of contagious caprine pleuropneumonia (CCPP)

Woubit, Salah; Lorenzon, Sophie; Peyraud, Armelle; Manso-Silván, Lucía; Thiaucourt, François

doi:10.1016/j.vetmic.2004.08.006

Cited by 68 publications

(58 citation statements)

References 15 publications

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“…capricolum MCAP0862 gene with 90% nucleotide identity. We therefore shortened the A51-F and A51-R primers (21-nucleotide primers designated ShA51-F and ShA51-R) to combine our PCR assay (simpA51) with that designed by Woubit et al (27). As illustrated in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…capripneumoniae type strains, 90% identity was obtained, underlining the high genetic relatedness of the two subspecies. We solved this problem by multiplexing the A51 PCR with the PCR described by Woubit et al (27) to exclude any non-M. capricolum subsp. capricolum isolates.…”

Section: Discussionmentioning

confidence: 99%

“…mycoides biotype Small Colony (20,28) and M. capricolum subsp. capripneumoniae (27) are available, but the accurate identification of the remaining taxa of the M. mycoides cluster is more problematic because of intertaxon cross-reactivity and high intrataxon variability (12). Two direct PCR assays currently are proposed in the literature for the specific identification of M. capricolum subsp.…”

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confidence: 99%

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Suppression-Subtractive Hybridization as a Strategy To Identify Taxon-Specific Sequences within the Mycoplasma mycoides Cluster: Design and Validation of an M. capricolum subsp. capricolum -Specific PCR Assay

Maigre¹,

Citti

Marenda

et al. 2008

J Clin Microbiol

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The phylogenetically related Mycoplasma capricolum subsp. capricolum and M. mycoides subsp. mycoides biotype Large Colony are two small-ruminant pathogens involved in contagious agalactia. Their respective contributions to clinical outbreaks are not well documented, because they are difficult to differentiate with the current diagnostic techniques. In order to identify DNA sequences specific to one taxon or the other, a suppression-subtractive hybridization approach was developed. DNA fragments resulting from the reciprocal subtraction of the type strains were used as probes on a panel of M. capricolum subsp. capricolum and M. mycoides subsp. mycoides biotype Large Colony strains to assess their intrataxon specificity. Due to a high intrataxon polymorphism and important cross-reactions between taxa, a single DNA fragment was shown to be specific for M. capricolum subsp. capricolum and to be present in all M. capricolum subsp. capricolum field isolates tested in this study. A PCR assay targeting the corresponding gene (simpA51) was designed that resulted in a 560-bp amplification only in M. capricolum subsp. capricolum and in M. capricolum subsp. capripneumoniae (the etiological agent of contagious caprine pleuropneumonia). simpA51 was further improved to generate a multiplex PCR (multA51) that allows the differentiation of M. capricolum subsp. capripneumoniae from M. capricolum subsp. capricolum. Both the simpA51 and multA51 assays accurately identify M. capricolum subsp. capricolum among other mycoplasmas, including all members of the M. mycoides cluster. simpA51 and multA51 PCRs are proposed as sensitive and robust tools for the specific identification of M. capricolum subsp. capricolum and M. capricolum subsp. capripneumoniae.The Mycoplasma genus is composed of wall-less bacteria with small genomes (0.58 to 1.35 Mb) and includes several species known to cause important diseases in humans and animals (24). Most of these species can be grown under laboratory conditions, although they require complex, undefined media and several days to several weeks of incubation because of their limited biosynthetic capacities (24). So far, mycoplasma identification and diagnosis have been based mainly on serological assays after cultivation, but recent ongoing efforts have been directed toward replacing these assays with more rapid and accurate molecular approaches based on the detection of specific DNA sequences.For the veterinary field, members of the so-called Mycoplasma mycoides cluster are of particular concern, because they are important ruminant pathogens that are responsible for economic losses worldwide (1, 28). This cluster includes six taxa, which are phylogenetically closely related based on their 16S rRNA gene sequences (21); present similar serological patterns associated with important intertaxon cross-reactivity (12); and, consequently, are difficult to differentiate. Four taxa contain pathogens for small ruminants, with three, namely, M. mycoides subsp. mycoides biotype Large Colony, M. mycoides subsp. capri,...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Suppression-Subtractive Hybridization as a Strategy To Identify Taxon-Specific Sequences within the Mycoplasma mycoides Cluster: Design and Validation of an M. capricolum subsp. capricolum -Specific PCR Assay

Maigre¹,

Citti

Marenda

et al. 2008

J Clin Microbiol

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“…Mccp and other members of the Mycoplasma mycoides cluster cross react in the serological test and share biochemical and genetic similarities, so biochemical and growth inhibition tests are not reliable and specific (Awan et al 2009;OIE 2009). The best and most accurate diagnostic method is molecular typing of Mccp (Woubit et al 2004). .…”

Section: Diagnostic Testsmentioning

confidence: 99%

Contagious caprine pleuropneumonia and its current picture in Pakistan: a review

Samiullah¹

2013

Vet. Med. - Czech

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Contagious caprine pleuropneumonia (CCPP) is caused by Mycoplasma capricolum subsp. capripneumoniae (Mccp) which belongs to the Mycoplasma mycoides cluster, a group of five closely related Mycoplasmas, pathogenic to ruminants. The true lesions of CCPP are restricted to the alveolar tissues of infected goats, which distinguish it from other respiratory diseases of small ruminants caused by members of the Mycoplasma mycoides cluster. The typical signs of CCPP are an accumulation of pleural fluid, unilateral hepatisation, adhesions, pleurisy and pleuropneumonia which clearly differentiate it from "MAKePS" syndrome caused by Mycoplasma mycoides subsp. capri (Mmc). The available literature on CCPP shows that so far in Pakistan, the true causative agent (Mccp) of this disease has only been isolated in the Pashin District of Balochistan and that the disease is more frequently confused with other respiratory diseases of goat caused by the Mycoplasma mycoides cluster. The lack of suitable techniques and extensive knowledge in the field is a big limitation for the isolation and characterisation of Mccp from prevailing CCPP-like cases in the goat population of Pakistan.

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“…capripneumoniae (MCCP), which occurs in many countries of Africa and Asia (Woubit et al, 2004). It is a disease of major economic relevance characterized by high morbidity and mortality; the mortality rate often approaches 100% in susceptible flocks (Rurangirwa et al, 1987).…”

Section: Introductionmentioning

confidence: 99%

Sensitive and rapid detection of Mycoplasma capricolum subsp. capripneumoniae by Loop-mediated isothermal amplification

He¹,

Zhang²,

Zhao³

et al. 2014

Afr. J. Biotechnol.

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A set of four specific primers was designed by targeting the H2 gene sequences of Mycoplasma capricolum subsp. capripneumoniae (MCCP). Using Bst DNA polymerase, the products were amplified for 60 min at 65°C in a simple water bath. Compared with a polymerase chain reaction (PCR) test that targets the H2 gene sequences of MCCP, the sensitivity of the loop-mediated isothermal amplification (LAMP) assay was higher (approximately 0.75 fg DNA per reaction). The LAMP products could be visualized by agar gel electrophoresis. There were no cross reactions with other strains in the Mycoplasma mycoides cluster, which indicates the high specificity of the LAMP procedure. The LAMP assay was able to detect MCCP in tissue.

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A specific PCR for the identification of Mycoplasma capricolum subsp. capripneumoniae, the causative agent of contagious caprine pleuropneumonia (CCPP)

Cited by 68 publications

References 15 publications

Suppression-Subtractive Hybridization as a Strategy To Identify Taxon-Specific Sequences within the Mycoplasma mycoides Cluster: Design and Validation of an M. capricolum subsp. capricolum -Specific PCR Assay

Suppression-Subtractive Hybridization as a Strategy To Identify Taxon-Specific Sequences within the Mycoplasma mycoides Cluster: Design and Validation of an M. capricolum subsp. capricolum -Specific PCR Assay

Contagious caprine pleuropneumonia and its current picture in Pakistan: a review

Sensitive and rapid detection of Mycoplasma capricolum subsp. capripneumoniae by Loop-mediated isothermal amplification

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