Enzyme-Linked Immunosorbent Assay and Microbiologic Culture for Diagnosis of Staphylococcus aureus Intramammary Infection in Cows

1996 ◽  
Vol 59 (1) ◽  
pp. 6-10 ◽  
Author(s):  
PAUL C. BARTLETT ◽  
RONALD J. ERSKINE ◽  
PATRICK GASTON ◽  
PHILIP M. SEARS ◽  
HENDRICUS WILHELMUS HOUDIJK
Keyword(s):  
Staphylococcus Aureus ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Relative Sensitivity ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infection Status ◽  
Intramammary Infection ◽  
Previous Infection ◽  
Current Infection

Recent reports have indicated that the relative sensitivity and specificity of the ELISA test for detection of intramammary infection of cows with Staphylococcus aureus is not as high as originally reported. It has been suggested that antibodies measured by enzyme-linked immunosorbent assay (ELISA) more closely reflect previous infection status rather than current infection status, and that the delay in antibody formation following infection and the persistence of antibodies after elimination of infection may be responsible for some of the discrepancy observed between ELISA and bacterial culture results conducted on the same milk sample. This study (n = 209 cows) was undertaken to determine if an ELISA for S. aureus intramammary infection more closely reflects previous infection status than it does current infection status, and to ascertain whether correction of this time-delay factor substantially improves calculated values of ELISA relative sensitivity and specificity. Receiver-operator curves were constructed to compare different time-related definitions of microbiologic culture results used for comparison with ELISA results. A greater degree of curvature in receiver-operator curves indicated that ELISA results did more closely reflect culture results performed on milk samples taken 1 and 3 weeks previously. Insignificant improvement in sensitivity and specificity occurred when the database was limited to cows (n = 140) with milk production greater than 13.6 kg/day. However, values of sensitivity were all less than or equal to 90%, and values of specificity were all less than 54%.

scholarly journals Indirect Enzyme Linked Immunosorbent Assay Sebagai Metode untuk Melacak Bruselosis pada Sapi Perah (INDIRECT ENZYME IMMUNOSORBENT ASSAY (I-ELISA) AS METHOD FOR DETECT BRUCELLOSIS IN DAIRY COW)

2019 ◽  
Vol 20 (1) ◽  
pp. 30
Author(s):  
Rinaldi Ghurafa ◽  
Denny Widaya Lukman ◽  
Hadri Latif
Keyword(s):  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Results ◽  
Kappa Value ◽  
Alternative Test ◽  
The Rose ◽  
Higher Sensitivity ◽  
Good Agreement

Brucellosis has become a zoonotic disease that received attention in efforts to prevent and eradicate strategic infectious animal diseases in Indonesia. Brucellosis can be detected early by the rose bengal test (RBT), followed by complement fixation test (CFT) and by enzyme linked immunosorbent assay (ELISA). The aims of this research was to study the indirect enzyme linked immunosorbent assay test (I-ELISA) as an alternative test for detecting brucellosis in dairy cattle. The method was used by conducting tests of RBT, CFT, I-ELISA and commercial I-ELISA to test brucellosis. The test results were calculated sensitivity and specificity, as well as analyzed by calculating the kappa value. The method was used by conducting tests of RBT, CFT, I-ELISA and commercial I-ELISA to test brucellosis. The test results were calculated for sensitivity and specificity, as well as analyzed by calculating the Kappa statistical value. The results of the sensitivity and specificity calculation showed that the indirect enzyme linked immunosorbent assay (I-ELISA) test developed a higher sensitivity (100%) compared to RBT test (93.75%) and commercial I-ELISA (93.75%). The developed I-ELISA specificity (74.68%) was still lower than RBT (89.87%), but higher than commercial I-ELISA (70.52%). The calculation of the statistical value of kappa RBT with CFT showed the kappa value 0.7120 which meaned it had a good agreement, commercial I-ELISA with CFT showed kappa value 0.6165 which meaned it had good suitability, whereas I-ELISA developed with CFT showed kappa value 0.4984 which meaned having a moderate agreement.In conclusion, the indirect enzyme linked immunosorbent assay (I-ELISA) which had been developed had low specificity, but the sensitivity was the highest compared to the commercial I-ELISA test and RBT, so this test was appropriate to be used as a screening test, especially in dairy cows movement into brucellosis-free areas or regions.

Integration of Liquid Chromatography with Immunoassay: An Approach Combining the Strengths of Both Methods

1994 ◽  
Vol 77 (5) ◽  
pp. 1275-1287 ◽  
Author(s):  
Petra M Krämer ◽  
Qing X Li ◽  
Bruce D Hammock
Keyword(s):  
Liquid Chromatography ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Uv Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Multiresidue Analysis ◽  
Selective Method ◽  
Separation Power ◽  
Immunochemical Detection ◽  
Very High

Abstract The integration of liquid chromatography (LC) with immunochemical detection combines the superior separation power of LC and the sensitivity and specificity of immunoassays. This approach is shown with 3 LC systems (Perkin-Elmer, C18 RP, 4.6 mm; Varian, C18 RP, 1 mm microbore; Michrom, C18 RP, 1 mm microbore) Integrated with an enzyme-linked immunosorbent assay (ELISA) selective for five 4-nitrophenols. The nitrophenols were separated with the 3 LC systems with isocratic runs of 15 to 20 min. Microbore LC separation showed a 10-20 times reduction in solvent amount compared to conventional separation. LC–immunoassay was about 8- to 10-fold more sensitive compared with LC with UV detection. Integrated LC–immunoassay proved to be a very selective method when 2-methylphenol was injected with an equimolar mixture of 2-amino-4-nitrophenol and 3-methyl-4-nrtrophenol; 2-methy I phenol does not crossreact with the serum used. Only 2 peaks could be seen in the detection, even when 2-methylphenol was present in very high amounts (3000 pmol). Further, the EUSA-LC detection proved to be selective and sensitive for complex matrixes. 2-Amlno-4-nitrophenol was clearly identified in spiked extracts of soil and plant, even when a very small amount (2.4 ng) was injected. Although LC–immunoassay is more labor intensive than LC with UV detection, it offers great advantages in multiresidue analysis and is generally applicable for peak confirmation.

Studies on the enzyme linked immunosorbent assay (ELISA) test forSchistosoma mansoniinfections

1978 ◽  
Vol 72 (3) ◽  
pp. 243-253 ◽  
Author(s):  
M. McLaren ◽  
C. C. Draper ◽  
J. M. Roberts ◽  
E. Minter-Goedbloed ◽  
G. S. Ligthart ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay

The effect of inoculating the bovine teat duct with small numbers of Staphylococcus aureus

1965 ◽  
Vol 32 (2) ◽  
pp. 171-179 ◽  
Author(s):  
F. H. S. Newbould ◽  
F. K. Neave
Keyword(s):  
Staphylococcus Aureus ◽  
Stainless Steel ◽  
Post Inoculation ◽  
Intramammary Infection ◽  
Colony Forming Units ◽  
Previous Infection ◽  
Glass Rods

SummaryFifty-seven inoculations of a strain of Staphylococcus aureus were made into the outer 4 mm of the teat ducts of 8 cows. The inocula ranged from 10 to 600 colony-forming units (CFU) and were made with a stainless steel instrument designed to overcome the disadvantages of glass rods and cotton swabs hitherto used.A single inoculation resulted in either no colonization of the duct, in temporary colonization for up to 7 days, or in colonization followed by intramammary infection.The recovery of the organisms depended on the size of the inoculum. When 70–100 CFU were placed in the teat duct no organisms were recovered from 12 of 24 quarters after the 1st post-inoculation milking. When 500–600 CFU were used, organisms were recovered from all of 31 quarters for at least 3 milkings, and from 61% for 6 milkings or more, in spite of dipping the teats in a strong disinfectant twice daily.Intramammary infection developed in 1 of 12 quarters (8%) inoculated in the teat duct with about 600 CFU when the animals were milked twice daily, and in 5 of 19 (23%) quarters if the 1st post-inoculation milking was omitted.There was no evidence of sensitization resulting from previous infection.

scholarly journals Desnutrição neonatal e produção de IFN-γ IL-12 e IL-10 por macrófagos/linfócitos: estudo da infecção celular, in vitro, por Staphylococcus aureus meticilina sensível e meticilina resistente

2012 ◽  
Vol 25 (5) ◽  
pp. 607-619 ◽  
Author(s):  
Thacianna Barreto da Costa ◽  
Natália Gomes de Morais ◽  
Thays Miranda de Almeida ◽  
Maiara Santos Severo ◽  
Célia Maria Machado Barbosa de Castro
Keyword(s):  
Staphylococcus Aureus ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ifn Γ

OBJETIVO: Avaliar a influência da desnutrição neonatal sobre a produção de Interferon gama, Interleucina-12 e Interleucina-10 em cultura de macrófagos alveolares e linfócitos infectados, in vitro, com Staphylococcus aureus sensível/resistente à meticilina. MÉTODOS: Ratos machos Wistar foram amamentados por mães cuja dieta, durante a lactação, continha 17% de proteína no grupo nutrido e 8% no grupo desnutrido. Após desmame, ambos os grupos receberam a dieta normoproteica. Os macrófagos foram obtidos após traqueostomia, através da coleta do lavado broncoalveolar. Para obtenção dos linfócitos, foi realizado o procedimento cirúrgico de punção cardíaca. Após o isolamento dos diferentes tipos celulares, procedeuse à realização dos estímulos com as cepas de estudo. A dosagem das citocinas foi realizada pelo método de Enzyme-Linked Immunosorbent Assay, a partir de amostras coletadas do sobrenadante das culturas após 24 horas de incubação. RESULTADOS: A desnutrição acarretou diminuição do crescimento ponderal, redução na produção de Interferon gama em cultura de macrófagos alveolares e linfócitos e diminuição na produção de Interleucina-12 em cultura de macrófagos alveolares. Apenas a produção de Interferon gama e Interleucina-10 em cultura de macrófagos alveolares apresentou diferença entre as cepas analisadas, em ambos os grupos estudados. CONCLUSÃO: O modelo de desnutrição neonatal produziu sequela no peso corporal e reduziu a produção de citocinas próinflamatórias (Interleucina-12 e Interferon gama), indicando que esse modelo de desnutrição pode comprometer a resolução de um processo infeccioso. A cepa de Staphylococcus aureus resistente à meticilina estimulou uma maior produção de Interferon gama e Interleucina-10 por macrófagos alveolares, o que sugeriu estimulação imunológica mais intensa, por essa cepa, nesse tipo celular especificamente.

scholarly journals Analysis ofLeishmaniamimetic neoglycoproteins for the cutaneous leishmaniasis diagnosis

Parasitology ◽  
2018 ◽  
Vol 145 (14) ◽  
pp. 1938-1948 ◽  
Author(s):  
Lígia Moraes Barizon de Souza ◽  
Vanete Thomaz Soccol ◽  
Ricardo Rasmussen Petterle ◽  
Michelle D. Bates ◽  
Paul A. Bates
Keyword(s):  
Cell Surface ◽  
Cutaneous Leishmaniasis ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Serum Levels ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
Cell Surfaces ◽  
Cell Markers ◽  
Antibody Titres

AbstractOligosaccharides are broadly present onLeishmaniacell surfaces. They can be useful for the leishmaniases diagnosis and also helpful in identifying new cell markers for the disease. The disaccharide Galα1-3Galβis the immunodominant saccharide inLeishmaniacell surface and is the unique non-reducing terminal glycosphingolipids structure recognized by anti-α-Gal. This study describes an enzyme-linked immunosorbent assay (ELISA) used to measure serum levels of anti-α-galactosyl (α-Gal) antibodies in patients with cutaneous leishmaniasis (CL). Optimal ELISA conditions were established and two neoglycoproteins (NGP) containing the Galα1-3Gal terminal fraction (Galα1-3Galβ1-4GlcNAc-HAS and Galα1-3Gal-HAS) and one Galα1-3Gal NGP analogue (Galα1-3Galβ1-3GlcNAc-HAS) were used as antigens. Means of anti-α-Gal antibody titres of CL patients were significantly higher (P< 0.05) than the healthy individuals for all NGPs tested. Sensitivity and specificity of all NGPs ranged from 62.2 to 78.4% and 58.3 to 96.7%, respectively. In conclusion, the NGPs can be used for CL diagnosis.

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