Detection of Mycoplasma mycoides subspecies mycoides by monoclonal antibody-based sandwich ELISA

Rodrı́guez, Francisco; Ball, H.J.; Finlay, D.; Campbell, D B; Mackie, D.P.

doi:10.1016/0378-1135(96)00018-1

Cited by 8 publications

(3 citation statements)

References 14 publications

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“…Immunohistochemistry using the antibody, 5E5, which previously has been shown to react with antigens on all mycoplasma of the mycoides cluster (30)(31)(32), demonstrated mycoplasma organisms in lesions from each of the three deceased rhebok (Fig. 1D, E, and F).…”

Section: Resultsmentioning

confidence: 80%

Systemic Disease in Vaal Rhebok ( Pelea capreolus ) Caused by Mycoplasmas in the Mycoides Cluster

Nicolas¹,

Stalis²,

Clippinger

et al. 2005

J Clin Microbiol

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In the winter of 2002, an outbreak of mycoplasma infection in Vaal rhebok (Pelea capreolus) originating from South Africa occurred 15 weeks after their arrival in San Diego, Calif. Three rhebok developed inappetence, weight loss, lethargy, signs related to pulmonary or arthral dysfunction, and sepsis. All three rhebok died or were euthanized. Primary postmortem findings were erosive tracheitis, pleuropneumonia, regional cellulitis, and necrotizing lymphadenitis. Mycoplasmas were detected in numerous tissues by electron microscopy, immunohistochemistry, and PCR. The three deceased rhebok were coinfected with ovine herpesvirus-2, and two animals additionally had a novel gammaherpesvirus. However, no lesions indicative of herpesvirus were seen microscopically in any animal. The rheboks' mycoplasmas were characterized at the level of the 16S rRNA gene, the 16S-23S intergenic spacer region, and the fructose biphosphate aldolase gene. Denaturing gradient gel electrophoresis was carried out to address the possibility of infection with multiple strains. Two of the deceased rhebok were infected with a single strain of Mycoplasma capricolum subsp. capricolum, and the third animal had a single, unique strain most closely related to Mycoplasma mycoides subsp. mycoides large-colony. A PCR survey of DNA samples from 46 other ruminant species demonstrated the presence of several species of mycoplasmas in the mycoides cluster, including a strain of M. capricolum subsp. capricolum identical to that found in two of the rhebok. These findings demonstrate the pervasiveness of mycoplasmas in the mycoides cluster in small ruminants and the potential for interspecies transmission and disease when different animal taxa come in contact.Mycoplasmas of the mycoides cluster are pathogens of ruminants and comprise six closely related species that are subdivided into two subgroups based on genetic similarity. The capricolum subgroup includes Mycoplasma capricolum subsp. capricolum, Mycoplasma capricolum subsp. capripneumoniae, and Mycoplasma subsp. bovine group 7 (BG7). The mycoides subgroup consists of Mycoplasma mycoides subsp. capri, Mycoplasma mycoides subsp. mycoides small-colony, and Mycoplasma mycoides subsp. mycoides large-colony (45). All six species of the mycoides cluster can cause respiratory, arthral, genitourinary, or mammary disease, although significant hostand strain-related variation in virulence exists. M. mycoides subsp. mycoides small-colony and M. capricolum subsp. capripneumoniae, the etiologies of contagious bovine pleuropneumonia (CBPP) and contagious caprine pleuropneumonia (CCPP), respectively, are the most virulent and typically induce fatal systemic disease in their hosts. Other members of the mycoides cluster, while pathogenic, do not usually cause life-threatening illness and, instead, establish protracted infections that result in low levels of chronic inflammation (8, 37).Infection of sheep and goats with mycoplasmas in the mycoides cluster occurs worldwide. The insidious nature of many of these infections al...

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Section: Resultsmentioning

confidence: 80%

Systemic Disease in Vaal Rhebok ( Pelea capreolus ) Caused by Mycoplasmas in the Mycoides Cluster

Nicolas¹,

Stalis²,

Clippinger

et al. 2005

J Clin Microbiol

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“…tnycoidees (LC and SC), and M. mycoides ssp. Capri and Myc+sma F38, respectively, when used in a sandwich enzyme-linked immunoabsorbent assay @LISA) format (RODRIGUEZ et al, 1996). Monoclonal antibody 5A10, which is specific for M. bovis and A{.…”

Section: Monoclonal Antibodies and Antiseramentioning

confidence: 99%

An Immunohistochemical Method of Detecting Mycoplasma Species Antigens by Use of Monoclonal Antibodies on Paraffin Sections of Pneumonic Bovine and Caprine Lungs

Rodrı́guez

Kennedy

Bryson

et al. 1996

Journal of Veterinary Medicine, Series B

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Summary Lung samples from pneumonic lesions in cattle and goats, naturally or experimentally infected with strains of the Mycoplasma mycoides cluster, were fixed in formalin and embedded in paraffin. An immunohistochemical technique using monoclonal or polyclonal antibodies was performed on tissue sections in order to detect Mycoplasma antigens. Four monoclonal antibodies (MAbs), one (2A3) raised against M. mycoides ssp. mycoides small colony (SC) and large colony (LC), two (1D3 and 5E5) against M. mycoides ssp. capri, and one (5A10) against M. bovis, were used. A range of polyclonal antibodies, raised to the individual subspecies of the M. mycoides cluster, and one to Pasteurella haemolytica, was also used. The MAb 2A3 showed positive immunostaining in lung sections from cattle and goats naturally and experimentally infected with M. mycoides ssp. mycoides SC and LC, but not with pneumonic lesions of cattle and goats due to other members of the M. mycoides cluster, M. bovis or Pasteurella spp. The MAb 1D3 showed immunostaining in lung sections from goats naturally and experimentally infected with M. mycoides ssp. capri, but again not with pneumonic lesions caused by other members of the M. mycoides cluster, M. bovis or Pasteurella spp. The MAb 5E5 immunoreacted in sections from pneumonic lesions from all animals infected with one of the three M. mycoides cluster subspecies used in the study, but not with M. bovis or Pasteurella infected tissue. Immunoreaction was mainly found in the cell debris around necrotic areas, as well as in macrophages, neutrophils and epithelial cells. The localization of antigens of the M. mycoides cluster using polyclonal antisera followed basically the same pattern as that obtained with the monoclonals. However, a wide cross reactivity was found between different antisera and relatively high background immunostaining was also seen, especially in necrotic areas. The results suggest that immunohistochemical methods using monoclonal antibodies are useful tools for the diagnosis and study of the pathogenesis of pneumonia caused by the Mycoplasmas of the M. mycoides cluster.

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“…Rodriguez et al (1996) used capture-ELISA for detection of M. mycoides subsp mycoides using monoclonal using monoclonal antibodies. The test was found to be specific and the sensitivity increased by overnight or 48 hr enrichment.…”

Section: Advances In Animal and Veterinary Sciencesmentioning

confidence: 99%

Detection of Mycoplasma equigenitalium from Equines by Capture- ELISA

Khurana¹

2015

Adv. Anim. Vet. Sci.

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| Mycoplasmas are smallest self-replicating prokaryotes and responsible for varied genital and respiratory disease conditions in animals as well as human beings. They assume greater importance due to their resistance for common antibiotics and ability to survive the temperature of cryopreservation during artificial insemination. Mycoplasma equigenitalium has been implicated in reproductive and genital problems of mares and stallions. The conventional cultural methods are quite cumbersome and time consuming. In this study detection rate by capture-ELISA (9.6%) was higher than cultural methods (4.8%) for diseased equines. Overall detection rate by capture-ELISA (3.6%) was also higher than cultural methods (1.8%). Therefore capture-ELISA becomes a simple, rapid and sensitive alternative tool for detection of M. equigenitalium.

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Detection of Mycoplasma mycoides subspecies mycoides by monoclonal antibody-based sandwich ELISA

Cited by 8 publications

References 14 publications

Systemic Disease in Vaal Rhebok ( Pelea capreolus ) Caused by Mycoplasmas in the Mycoides Cluster

Systemic Disease in Vaal Rhebok ( Pelea capreolus ) Caused by Mycoplasmas in the Mycoides Cluster

An Immunohistochemical Method of Detecting Mycoplasma Species Antigens by Use of Monoclonal Antibodies on Paraffin Sections of Pneumonic Bovine and Caprine Lungs

Detection of Mycoplasma equigenitalium from Equines by Capture- ELISA

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