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.

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%.

Novel Method for ELISA Traces Pipetting System by Air Displacement Pipetting

2011 ◽  
Vol 103 ◽  
pp. 252-256 ◽  
Author(s):  
Lian Qing Zhu ◽  
Hong Li ◽  
Yun Xiao Na ◽  
Yang Kuan Guo ◽  
Ming Li Dong
Keyword(s):  
Real Time ◽  
Immunosorbent Assay ◽  
Pressure Sensor ◽  
Applied Pressure ◽  
Elisa Test ◽  
Liquid Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Results ◽  
The Real ◽  
Novel Method

In the Enzyme-linked immunosorbent assay (ELISA) test, many steps need traces pipetting. The ELISA test results will be different when we use different pipetting ways. Our traces pipetting system is based on the air displacement pipetting principle, comparable to the functioning of hand pipettes. It is applied pressure sensor to realize pressure-based liquid level detection (pLLD) and aspiration monitoring. The monitored system can distinguish the following situations: (1) a correct aspiration; (2) cup empty; (3) tip-blocked; (4) bubbles. Using the air displacement principle into traces pipetting can avoid contamination or dilution by system liquids, and problems with corroded tubing, pumps, etc. It applied pressure sensor to realize pLLD and aspiration monitoring. The results of the real-time monitor module on air displacement pipetting show that the traces pipetting system can agilely distinguish the different liquid pipetting situations. The method of air displacement pipetting offered an effective way for ELISA traces pipetting system.

scholarly journals Development of a Rapid Diagnostic Kit That Uses an Immunochromatographic Device To Detect Antibodies in Human Sparganosis

2014 ◽  
Vol 21 (9) ◽  
pp. 1360-1363 ◽  
Author(s):  
Hiroshi Yamasaki ◽  
Takeshi Nakamura ◽  
Pewpan M. Intapan ◽  
Wanchai Maleewong ◽  
Yasuyuki Morishima ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Laboratory Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Sparganosis ◽  
Higher Sensitivity

ABSTRACTA diagnostic kit using an immunochromatographic device was developed to replace the time-consuming immunodiagnostic methods for human sparganosis. The kit was found to be faster and easier to use than an enzyme-linked immunosorbent assay (ELISA) and showed higher sensitivity and specificity. It will be useful for the laboratory diagnosis of hospitalized cases of sparganosis.

scholarly journals Evaluation of Enzyme-Linked Immunosorbent Assay for Diagnosis of Post-Kala-Azar Dermal Leishmaniasis with Crude or Recombinant k39 Antigen

2002 ◽  
Vol 9 (2) ◽  
pp. 370-373 ◽  
Author(s):  
P. Salotra ◽  
G. Sreenivas ◽  
A. A. Nasim ◽  
B. V. Subba Raju ◽  
V. Ramesh
Keyword(s):  
Immunosorbent Assay ◽  
Leishmania Donovani ◽  
Skin Lesions ◽  
Elisa Test ◽  
Recombinant Antigen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Invasive Test ◽  
Kala Azar ◽  
Axenic Amastigotes ◽  
Higher Sensitivity

ABSTRACT The diagnosis of post-kala-azar dermal leishmaniasis (PKDL), a dermatosis that provides the only known reservoir for the parasite Leishmania donovani in India, remains a problem. Timely recognition and treatment of PKDL would contribute significantly to the control of kala-azar. We evaluated here the potential of the enzyme-linked immunosorbent assay (ELISA) as a diagnostic tool for PKDL. Antigen prepared from promastigotes and axenic amastigotes with parasite isolates that were derived from skin lesions of a PKDL patient gave sensitivities of 86.36 and 92%, respectively, in the 88 PKDL cases examined. The specificity of the ELISA test was examined by testing groups of patients with other skin disorders (leprosy and vitiligo) or coendemic infections (malaria and tuberculosis), as well as healthy controls from areas where this disease is endemic or is not endemic. A false-positive reaction was obtained in 14 of 144 (9.8%) of the controls with the promastigote antigen and in 14 of 145 (9.7%) of the controls with the amastigote antigen. Evaluation of the serodiagnostic potential of recombinant k39 by ELISA revealed a higher sensitivity (94.5%) and specificity (93.7%) compared to the other two antigens used. The data demonstrate that ELISA with crude or recombinant antigen k39 provides a relatively simple and less-invasive test for the reliable diagnosis of PKDL.

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

scholarly journals Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

2020 ◽  
Vol 8 (8) ◽  
pp. 1235 ◽  
Author(s):  
Mareike Stellfeld ◽  
Claudia Gerlach ◽  
Ina-Gabriele Richter ◽  
Peter Miethe ◽  
Dominika Fahlbusch ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Roc Curves ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Domestic Ruminants ◽  
Diagnostic Potential ◽  
Assay Performance ◽  
Higher Sensitivity

Coxiella burnetii is the causative agent of Q fever, a zoonosis infecting domestic ruminants and humans. Currently used routine diagnostic tools offer limited sensitivity and specificity and symptomless infected animals may be missed. Therefore, diagnostic tools of higher sensitivity and specificity must be developed. For this purpose, the C. burnetii outer membrane protein Com1 was cloned and expressed in Escherichia coli. The His-tagged recombinant protein was purified and used in an indirect enzyme-linked immunosorbent assay (ELISA). Assay performance was tested with more than 400 positive and negative sera from sheep, goats and cattle from 36 locations. Calculation of sensitivity and specificity was undertaken using receiver operating characteristic (ROC) curves. The sensitivities and specificities for sheep were 85% and 68% (optical density at 450nm, OD450 cut-off value 0.32), for goats 94% and 77% (OD450 cut-off value 0.23) and for cattle 71% and 70% (OD450 cut-off value 0.18), respectively. These results correspond to excellent, outstanding and acceptable discrimination of positive and negative sera. In summary, recombinant Com1 can provide a basis for more sensitive and specific diagnostic tools in veterinary medicine.

scholarly journals Secondary Serological Response of Patients with Chronic Hepatosplenic Suppurative Brucellosis

2006 ◽  
Vol 13 (11) ◽  
pp. 1190-1196 ◽  
Author(s):  
R. Díaz ◽  
J. Ariza ◽  
I. Alberola ◽  
A. Casanova ◽  
M. F. Rubio
Keyword(s):  
Immunological Response ◽  
Lateral Flow ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Response ◽  
Test Results ◽  
Coombs Test ◽  
Igg Antibodies ◽  
Additional Information ◽  
The Rose

ABSTRACT Chronic hepatosplenic suppurative brucellosis (CHSB) is a local reactivation of a previous brucellosis, coursing with an immunoglobulin G (IgG) and IgA secondary immunological response. The observation of two cases of CHSB with an apparent IgM response gave rise to a detailed serological study of three of our patients. We studied the first sample from all three patients and successive samples from two of them. In cases 1 and 2, we found samples with positive IgM lateral flow and IgM enzyme-linked immunosorbent assay results concomitantly with rheumatoid factor (RF); after absorption with anti-RF serum, these results were rendered negative. In patients 2 and 3 the diagnosis of brucellosis was delayed, because none of the test results were initially very significant. However, a clear seroconversion of IgG antibodies was observed in subsequent months; titers of the Brucellacapt and Coombs tests increased in similar ways, although Brucellacapt decreased more rapidly than Coombs, which persisted at high titers for years. In patient 3 a relapse was observed in the fourth year of follow-up, detected by Coombs and also by IgG lateral flow and counterimmunoelectrophoresis (CIEP), although not by the rose bengal, agglutination, or Brucellacapt tests. Serological changes in CHSB may sometimes be mild and are detected mainly by the Coombs test. Brucellacapt does not offer additional information, although IgG lateral flow and CIEP may be of some use. Careful surveillance of titer changes in the Coombs test is the best marker of infection activity. As the disease progresses, an intense IgG response may develop and RF sometimes appears, simulating an IgM response.

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