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

Opitz, H. M.; Cyr, M J

doi:10.2307/1590636

Cited by 18 publications

(7 citation statements)

References 7 publications

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“…The obtained results of both whole antigen or sonicated antigen gave similar results and good binding affinity for the surface of the microtiter plates polystyrene wells, which go parallel to Optiz and Cyr 12 results that recorded strong adsorbing affinity to the surface of the microtiter plates polystyrene wells at prepared the myco-ELISA antigen from disrupted M.gallisepticum or whole bacterial cell.…”

Section: Discussionsupporting

confidence: 77%

Preparation of ELISA and Lateral Flow Kits for rapid Diagnosis of Mycoplasma gallisepticum in Poultry

El-Yazeed

Al-Atfeehy²,

Abotaleb³

et al. 2020

Sci Rep

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Avian mycoplasmas were mainly the cause of poultry industry economic losses; reduced meat and egg production and increases the antibiotic treatment cost. Mycoplasma gallisepticum (MG) infection is designated as infectious sinusitis of turkeys and chronic respiratory disease of chickens (gasping, depression, semi closed eyes, infraorbital sinuses edema and decrease in egg production). This study aimed to prepare, evaluate and Compare in-house ELISA kits and lateral flow assay (LFA) from a local strain of MG with commercial ELISA kits and PCR consequently. A total of 54 samples (27 tracheal swabs, 10 trachea and 17 lung) and 50 serum samples collected from birds suffering from chronic respiratory disease were tested by prepared in-house ELISA, commercial ELISA kits, PCR and LFA; a high correlation coefficient between in-house ELISA using whole antigen or sonicated antigen and commercial kit was recorded. Lateral Flow assay (LFA) performance indicate a low sensitivity (77.5%) but maintain a high specificity (92%) compared to PCR. The in-house ELISA kits and LFA prepared could be used as a fast diagnostic technique for detection of MG in Egypt. According to the available knowledge the prepared LFA for diagnosis of MG infection in chickens was developed for the first time in Egypt.

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Section: Discussionsupporting

confidence: 77%

Preparation of ELISA and Lateral Flow Kits for rapid Diagnosis of Mycoplasma gallisepticum in Poultry

El-Yazeed

Al-Atfeehy²,

Abotaleb³

et al. 2020

Sci Rep

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“…However cross reactions were demonstrated by others (Opitz & Cyr, 1986;Higgins & Whithear, 1986;Bradley et al, 1988).…”

Section: Discussionmentioning

confidence: 89%

Effect of duration of incubation ofMycoplasma gallisepticumcultures on the sensitivity and specificity of antigens for ELISA and microimmunofluorescence tests

1993

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SUMMARYIn the production of Mycoplasma gallisepticum (Mg) antigens for ELISA and microimmunofluorescence (MIF) tests, two strains of Mg, S6 and PG31, were grown in broth culture and harvested at intervals from 18 to 138 h. The effect of duration of incubation of culture on antigen sensitivity and specificity was assessed using homologous sera (against Mg S6), and sera prepared against Mycoplasma synoviae (Ms), and against mycoplasma media. It was found, in both Mg S6 and PG31 ELISAs, that in the period 18 to 70 h of incubation, sensitivity declined with homologous sera, but little thereafter. A more rapid and greater decline occurred with PG31 antigen than S6. The antigens showed a lack of specificity since antisera to Ms gave positive reactions. These were not influenced by duration of incubation of Mg culture. In the Mg S6 ELISA lack of specificity was also shown by positive reactions with sera against media components. These increased in the first 42 h of cultivation. In the MIF test there was no loss of sensitivity with Mg S6 antigen with increasing duration of incubation of culture but it occurred with PG31 antigen. The non-specific reactions with antisera to Ms increased with increased duration of incubation of both strains of Mg. This work demonstrates that the most sensitive antigens are produced by harvesting organisms early in culture, although this does not eliminate the Ms cross reactions.

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“…Several studies have reported increased sensitivity of M. synoviae ELISAs using different antigen preparations (Opitz et al, 1983;Patten et al, 1984;Higgins & Whithear, 1986;Opitz & Cyr, 1986;Avakian & Kleven, 1990;Gurevich et al, 1995) ; however, specificity continues to be a problem associated with M. synoviae ELIS As.…”

Section: Discussionmentioning

confidence: 99%

“…A number of earlier studies of M . synouiae ELISAs used crude membrane preparations (Opitz et al, 1983;Patten et al, 1984;Higgins & Whithear, 1986), produced by either osmotic lysis or sonication of whole cells, or Triton X-100-solubilized membrane proteins (Opitz & Cyr, 1986) of M . synouiae, as antigens.…”

Section: Introductionmentioning

confidence: 99%

Mycoplasma synoviae surface protein MSPB as a recombinant antigen in an indirect ELISA

Noormohammadi¹,

Markham²,

Markham³

et al. 1999

Microbiology

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~ ~~~Mycoplasma synoviae is a poultry pathogen causing respiratory disease and synovitis. A number of serological assays have been developed for diagnosis of M. synoviae infection; however, they lack sensitivity and/or are prone to false-positive reactions. Using a combination of PCR and expression cloning, four overlapping regions (regions 1 4 ) of the surface antigen MSPB of M.synoviae WVU-1853 were expressed in a bacterial expression system.lmmunostaining of the resultant polypeptides with chicken sera raised against different M. synoviae strains, or Mycoplasma gallisepticum S6, suggested that region 4 contained a highly antigenic and species-specific domain (amino acids 178-213) of MSPB. A fusion protein of region 4 was expressed in the pMAL expression system and purified from cold-osmotic-shock fluids of Escherichia coli cells for use in an indirect ELISA. The potential of the purified antigen for detection of M. synoviae antibodies was assessed with sera obtained from chickens experimentally infected with different strains of M. synoviae or M. gallisepticum, or from uninoculated chickens. All the sera from M. synoviaeinoculated chickens provided higher absorbance values than those from M. gallisepticum-inoculated or uninoculated chickens. Chickens inoculated with M. synoviae 86079nNS had detectable increases of serum anti-MSPB immunoglobulins a t day 7 after inoculation. These studies have identified the most antigenic region of one of the major species-specific surface proteins of M. synoviae, and shown the potential of this protein as a serodiagnostic reagent.

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

Abstract: Triton X-100-solubilized Mycoplasma gallisepticum and M. synoviae ELISA antigens were found to be more specific and sensitive than six other antigens at a concentration of 250 ng protein/0.1 ml per microtiter plate well.

Cited by 18 publications

References 7 publications

Preparation of ELISA and Lateral Flow Kits for rapid Diagnosis of Mycoplasma gallisepticum in Poultry

Preparation of ELISA and Lateral Flow Kits for rapid Diagnosis of Mycoplasma gallisepticum in Poultry

Effect of duration of incubation ofMycoplasma gallisepticumcultures on the sensitivity and specificity of antigens for ELISA and microimmunofluorescence tests

Mycoplasma synoviae surface protein MSPB as a recombinant antigen in an indirect ELISA

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