A PCR-based Test for the Specific Identification of <i>Mycoplasma Mycoides</i> Subspecies <i>mycoides</i> SC

Bashiruddin, J. B.; Taylor, Tamara; Gould, Alan R.

doi:10.1177/104063879400600405

Cited by 83 publications

(55 citation statements)

References 7 publications

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“…The oligonucleotide primer pair MMMSCO5-7 and MMMSCO5-6 can therefore be used in a confirmatory PCR method for the genetic identification of M. mycoides subsp. mycoides SC, together with previously established methods (6,13,24). It must be noted that no DNA sequence with similarity to that of lppQ has been found in other species of the class Mollicutes, based on currently accessible DNA sequences in databases.…”

Section: Discussionmentioning

confidence: 99%

Antigenic and Genetic Characterization of Lipoprotein LppQ fromMycoplasma mycoidessubsp.mycoidesSC

Abdo

Nicolet

Frey

2000

Clin Diagn Lab Immunol

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Lipoprotein LppQ, a predominant 48-kDa antigen, and its corresponding gene, lppQ, were characterized in Mycoplasma mycoides subsp. mycoides SC, the etiological agent of contagious bovine pleuropneumonia. The lppQ gene is specific to M. mycoides subsp. mycoides SC and was found in the type strain and in field strains isolated in Europe, Africa, and Australia, as well as in vaccinal strains. LppQ is encoded as a precursor with a consensus sequence for prokaryotic signal peptidase II and a lipid attachment site. The leader sequence shows significant prominent transmembrane helix structure with a predicted outside-to-inside helix formation capacity. The N-terminal domain of the mature LppQ was shown to be surface exposed. It induced a strong, specific, early, and persistent immune response in naturally and experimentally infected animals. The C-terminal domain of LppQ possesses an integral membrane structure built up of repeated units, rich in hydrophobic and aromatic amino acids, which have a pore formation potential. A recombinant peptide representing the N-terminal domain of LppQ was obtained by site-directed mutagenesis of nine Mycoplasma-specific TGA (Trp) codons into universal TGG (Trp) codons and expression in Escherichia coli hosts. It was used for serodetection of cattle infected with M. mycoides subsp. mycoides SC, in which it was detected postinfection for significantly longer than conventional serological test reactions.Mycoplasma mycoides subsp. mycoides small-colony type (SC) is the etiological agent of contagious bovine pleuropneumonia (CBPP), a highly contagious disease which represents a major threat to raising cattle, particularly in Africa. CBPP is also a problem in other parts of the world, including some European countries, where the disease suddenly reemerged 2 decades ago. CBPP is a disease of major economic concern in the affected countries, not only due to the morbidity and mortality but also due to restrictions on cattle trade imposed by international regulations. Hence, control of the disease is a priority for countries in which it is endemic, in order to eradicate the disease as quickly as possible after outbreaks and to avoid its spreading, as well as for countries which are free of CBPP, in order to keep that status. The main problem in eradication is the frequent occurrence of subacute or asymptomatic infections and the persistence of chronic carriers after the clinical phase. Serological analysis is the most important diagnostic tool for the control of CBPP, but it is significantly hampered by the relatively low sensitivity and specificity of the methods. The complement fixation test (CFT), which is currently the official and most widely used serodiagnostic test, has been shown to be relatively sensitive in the acute phase of the disease, but it levels off rather quickly and is insensitive 3 months after infection (1, 29). In contrast to CFT, immunoglobulin G (IgG) and IgA reactions to many antigens of M. mycoides subsp. mycoides SC are persistent for several months, as shown by immun...

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Section: Discussionmentioning

confidence: 99%

Antigenic and Genetic Characterization of Lipoprotein LppQ fromMycoplasma mycoidessubsp.mycoidesSC

Abdo

Nicolet

Frey

2000

Clin Diagn Lab Immunol

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“…Whilst serological classification and differentiation of cluster members is difficult, it has been shown that they can be distinguished by DNAbased diagnostic methods. The CAP-21 genomic region was used by Taylor et al (1992) to design a DNA probe identifying M. mycoides subspecies and also a PCR assay for identification of the SC type (Bashiruddin et al, 1994). Hotzel et al (1996) developed a PCR system for differentiation of all cluster groups which exploited nucleotide sequence variations in the same region.…”

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confidence: 99%

Comparison of the 16S-23S rRNA intergenic spacer regions among strains of the Mycoplasma mycoides cluster, and reassessment of the taxonomic position of Mycoplasma sp. bovine group 7.

Harasawa¹,

Hotzel²,

Sachse³

2000

International Journal of Systematic and Evolutionary Microbiology

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Nucleotide sequence analysis of the 16S-23S rRNA intergenic spacer regions of six type or reference strains belonging to the Mycoplasma mycoides cluster and of Mycoplasma putrefaciens suggested the presence of two subclusters. One subcluster comprised M. mycoides subsp. mycoides small colony (SC) type, M. mycoides subsp. mycoides large colony (LC) type and M. mycoides subsp. capri, whereas the second subcluster comprised Mycoplasma capricolum subsp. capricolum, M. capricolum subsp. capripneumoniae and Mycoplasma sp. bovine group 7. The type strains from M. mycoides subsp. mycoides SC and M. mycoides subsp. capri had identical spacer sequences. The existence of two subclusters was supported by predicted secondary structures of the analysed region. The nucleotide variations in the loop domains of the secondary structures might be a useful genetic marker to distinguish between the two subclusters. The secondary structure differences delineated the differences between the two subclusters more clearly than the nucleotide sequence alignments, which only showed a small number of differences, and some of these were common to both clusters. The data also provided evidence in favour of a reclassification of Mycoplasma sp. bovine group 7 as another subspecies of M. capricolum.

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“…The use of molecular tests directly on clinical specimens may improve the sensitivity of testing as the tests are not reliant on containing viable bacteria. However, the use of specific PCRs, such as the M. conjunctivae PCR and the Mycoplasma ''mycoides cluster'' and Mycoplasma ''mycoides subcluster'' PCRs coupled with restriction fragment length polymorphisms (Bashiruddin et al, 1994) will only detect and identify the specific Mycoplasma species the investigator is targeting and requires the use of several different tests. The recently introduced PCR/DGGE FIGURE 3.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Mycoplasma Isolated From an Ibex (Capra Ibex) Suffering From Keratoconjunctivitis in Northern Italy

Giangaspero

Orusa

Nicholas

et al. 2010

Journal of Wildlife Diseases

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ABSTRACT:In 2005 a Mycoplasma species was isolated from ocular-conjunctival swabs from an adult male Alpine ibex (Capra ibex) from the Valle d'Aosta Region, Northern Italy. The animal suffered from bilateral ocular discharge with diffuse inflammation, severe corneal involvement of the left eye and mild corneal opacity of the right eye. Histologic examination revealed a keratoconjunctivitis characterized by lymphocytic and plasmacellular infiltration. Laboratory investigations of the isolate included culture, transmission electron microscopy, PCR, and denaturing gradient gel electrophoresis, as well as DNA sequencing of the 16S rDNA gene. These tests identified the isolate as Mycoplasma mycoides subsp. capri large-colony serovar, an organism that has occasionally been associated with keratoconjunctivitis in goats. For a correct diagnosis, it is necessary to carry out laboratory investigations, as clinical cases of keratoconjunctivitis in wild ruminants are not always ascribable to Mycoplasma conjunctivae.

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A PCR-based Test for the Specific Identification of Mycoplasma Mycoides Subspecies mycoides SC

Cited by 83 publications

References 7 publications

Antigenic and Genetic Characterization of Lipoprotein LppQ fromMycoplasma mycoidessubsp.mycoidesSC

Antigenic and Genetic Characterization of Lipoprotein LppQ fromMycoplasma mycoidessubsp.mycoidesSC

Comparison of the 16S-23S rRNA intergenic spacer regions among strains of the Mycoplasma mycoides cluster, and reassessment of the taxonomic position of Mycoplasma sp. bovine group 7.

Characterization of Mycoplasma Isolated From an Ibex (Capra Ibex) Suffering From Keratoconjunctivitis in Northern Italy

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