Tween 20 soluble proteins of Mycoplasma hyopneumoniae as antigen for an enzyme linked immunosorbent assay

Nicolet, Jacques; Paroz, P; Bruggmann, S.

doi:10.1016/s0034-5288(18)32632-8

Cited by 51 publications

(16 citation statements)

References 11 publications

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“…Our results agree with Nicholet et al (4) in that Tween 20 treatment results in a more specific antigen than SDS treatment. Triton X-100 treatment solubilized 69.8% and 76.4% of the protein of the MG and MS WC antigens, respectively, which is a greater yield than that obtained by the method used earlier (5).…”

Section: Methodssupporting

confidence: 93%

“…WC MG and MS antigens were treated by various methods in attempts to improve antigen quality: WC antigen (1); disrupted WC (DWC) antigen (2); sonication of WC and DWC antigens; disrupted and fractionated antigen (3); Salsbury-stained plate antigen; WC, DWC, and yolk-sac-propagated antigens purified by discontinuous sucrose gradient; and antigens solubilized by 1% detergent of sodium dodecyl sulfate (SDS) (1), N-octyl glucoside (OG), Tween 20 (4), or Triton X-100. WC MG and MS antigens were treated by various methods in attempts to improve antigen quality: WC antigen (1); disrupted WC (DWC) antigen (2); sonication of WC and DWC antigens; disrupted and fractionated antigen (3); Salsbury-stained plate antigen; WC, DWC, and yolk-sac-propagated antigens purified by discontinuous sucrose gradient; and antigens solubilized by 1% detergent of sodium dodecyl sulfate (SDS) (1), N-octyl glucoside (OG), Tween 20 (4), or Triton X-100.…”

Section: Methodsmentioning

confidence: 99%

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Triton X-100-Solubilized Mycoplasma gallisepticum and M. synoviae ELISA Antigens

Opitz¹,

Cyr²

1986

Avian Diseases

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Section: Methodssupporting

confidence: 93%

Section: Methodsmentioning

confidence: 99%

Triton X-100-Solubilized Mycoplasma gallisepticum and M. synoviae ELISA Antigens

Opitz¹,

Cyr²

1986

Avian Diseases

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“…ELISA is considered the most useful test for detection of antibodies to M. hyopneumoniae. Many attempts have been made to purify the specific antigen from M. hyopneumoniae (1,15). However, it is very difficult to remove cross-reactivities against other swine mycoplasmas, especially against Mycoplasma flocculare.…”

Section: Discussionmentioning

confidence: 99%

“…A reliable diagnosis of M. hyopneumoniae infection is necessary for control of MPS. Several serological tests, such as the complement fixation test (14), indirect hemagglutination test (3), and enzyme-linked immunosorbent assay (ELISA) (1,2,15), have been developed to demonstrate antibodies to M. hyopneumoniae in the sera of diseased pigs, but these techniques were not completely satisfactory because of cross-reaction with other swine mycoplasmas. The double-sandwich ELISA method with a monoclonal antibody that recognized the 46-kDa surface antigen (P46) of M. hyopneumoniae was used successfully for the diagnosis of MPS (12).…”

mentioning

confidence: 99%

Recombinant 46-kilodalton surface antigen (P46) of Mycoplasma hyopneumoniae expressed in Escherichia coli can be used for early specific diagnosis of mycoplasmal pneumonia of swine by enzyme-linked immunosorbent assay

et al. 1995

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The 46-kDa surface antigen (P46) is the early and species-specific immunogenic protein of Mycoplasma hyopneumoniae. Three TGA codons encoding tryptophan in the P46 gene were replaced with TGG by an in vitro mutagenesis technique. The mutated P46 gene was expressed in Escherichia coli by using the chelating peptide tag system. The purified recombinant P46 was successfully used in an enzyme-linked immunosorbent assay for detection of antibodies against M. hyopneumoniae in swine serum. It did not cross-react with sera from swine infected with Mycoplasma flocculate, Mycoplasma hyorhinis, or Mycoplasma hyosynoviae. With this method, mycoplasmal pneumonia of swine was detectable within 2 weeks after infection.

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“…Several workers used purified Tween 20 extracts of M. hyopneumoniae and found superior specificity (Nicolet and Paroz 1980;Kazama et al 1989). These improved ELISA antigens have been described as suitable for the practical serodiagnosis of mycoplasmal pneumonia of swine (Yagihashi et al, 1993) and are available in a commercial M. hyopneumoniae ELISA test kit.…”

Section: Discussionmentioning

confidence: 99%

The pathogenesis of respiratory infection of lambs by mycoplasmas

Niang

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Conclusions M. ovipneumoniae expresses a polysaccharide capsule with variable size expression in different cultures. Strain heterogeneity could be further indicated by variable reactivity of this capsular material with several different lectins. Morphological observations suggest that this capsule facilitates adherence of the organism to ciliated epithelium. Clinical Relevance Strain variation associated with this capsule and its possible association with attachment to cilia may provide a versatile interaction of M. ovipneumoniae with its environment; thus, a long term colonization of the host by the organism. different isolates. Light and electron microscopic examinations of infected tracheal rings revealed disorganization and sloughing of the epithelium, and association of mycoplasmas only with the cilia. Following repeated in vitro passages, the organisms had reduced ability to inhibit ciliary activity which correlated with decreased hydrogen peroxide production. Addition of catalase to the organ cultures delayed loss of ciliary activity. Conclusions M. ovipneumoniae induced ciliostasis in ovine tracheal ring explants which correlated with hydrogen peroxide production. M. ovipneumoniae is an extracellular pathogen that colonizes the ciliated epithelium. Clinical relevance M. ovipneumoniae colonization of respiratory epithelial cells contributes to the pathogenesis of sheep respiratory disease.

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Tween 20 soluble proteins of Mycoplasma hyopneumoniae as antigen for an enzyme linked immunosorbent assay

Cited by 51 publications

References 11 publications

Triton X-100-Solubilized Mycoplasma gallisepticum and M. synoviae ELISA Antigens

Triton X-100-Solubilized Mycoplasma gallisepticum and M. synoviae ELISA Antigens

Recombinant 46-kilodalton surface antigen (P46) of Mycoplasma hyopneumoniae expressed in Escherichia coli can be used for early specific diagnosis of mycoplasmal pneumonia of swine by enzyme-linked immunosorbent assay

The pathogenesis of respiratory infection of lambs by mycoplasmas

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