Immunodiagnostic monoclonal antibody-based sandwich ELISA of fasciolosis by detection ofFasciola giganticacirculating fatty acid binding protein

Parasitology ◽  
2016 ◽  
Vol 143 (11) ◽  
pp. 1369-1381 ◽  
Author(s):  
PANAT ANURACPREEDA ◽  
RUNGLAWAN CHAWENGKIRTTIKUL ◽  
PRASERT SOBHON
Keyword(s):  
Monoclonal Antibody ◽  
Fatty Acid ◽  
Fatty Acid Binding Protein ◽  
Binding Protein ◽  
Sandwich Elisa ◽  
Parasite Burden ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fatty Acid Binding ◽  
Acid Binding Protein ◽  
Low Sensitivity

SUMMARYUp to now, parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Hence, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden. In the present study, a monoclonal antibody (MoAb) against recombinantFasciola giganticafatty acid binding protein (rFgFABP) has been produced. As well, a reliable sandwich enzyme-linked immunosorbent assay (sandwich ELISA) has been developed for the detection of circulating FABP in the sera of mice experimentally and cattle naturally infected withF. gigantica. MoAb 3A3 and biotinylated rabbit anti-recombinant FABP antibody were selected due to their high reactivities and specificities. The lower detection limit of sandwich ELISA was 5 pg mL−1, and no cross-reaction with other parasite antigens was observed. This assay could detectF. giganticainfection from day 1 post infection. In experimental mice, the sensitivity, specificity and accuracy of this assay were 93·3, 100 and 98·2%, while in natural cattle they were 96·7, 100 and 99·1%. Hence, this sandwich ELISA method showed high efficiencies and precisions for diagnosis of fasciolosis byF. gigantica.

Diagnostic potential ofFasciola gigantica-derived 14.5 kDa fatty acid binding protein in the immunodiagnosis of bubaline fascioliasis

2012 ◽  
Vol 87 (2) ◽  
pp. 147-153 ◽  
Author(s):  
G. Allam ◽  
I.R. Bauomy ◽  
Z.M. Hemyeda ◽  
T.M. Diab ◽  
T.F. Sakran
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Binding Protein ◽  
Binding Protein ◽  
Sandwich Elisa ◽  
Fasciola Gigantica ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Efficacy ◽  
Fatty Acid Binding ◽  
Acid Binding Protein ◽  
Diagnostic Potential

AbstractThe 14.5 kDa fatty acid binding protein (FABP) was isolated from the crude extract of adultFasciola giganticaworms. Polyclonal anti-FABP IgG was generated in rabbits immunized with prepared FABP antigen. Sandwich enzyme-linked immunosorbent assay (ELISA) was applied to detect coproantigen in stools and circulatingFasciolaantigen (CA) in sera of 126 water buffaloes by using purified and horseradish peroxidase (HRP)-conjugated anti-FABP IgG. Sandwich ELISA sensitivity was 96.97% and 94.95%; while specificity was 94.12% and 82.35% for coproantigen and CA detection, respectively. However, sensitivity and specificity of the Kato–Katz technique was 73.74% and 100%, respectively. The diagnostic efficacy of sandwich ELISA was 96.55% and 93.1% for coproantigen and CA detection, respectively. In contrast, the diagnostic efficacy of the Kato–Katz technique was 77.59%. In conclusion, these results demonstrate that the purified 14.5 kDa FABP provides a more suitable antigen for immunodiagnosis of early and current bubaline fascioliasis by using sandwich ELISA.

scholarly journals Epidermal Fatty Acid-Binding Protein: A Novel Marker in the Diagnosis of Dry Eye Disease in Sjögren Syndrome

2018 ◽  
Vol 19 (11) ◽  
pp. 3463 ◽  
Author(s):  
Megumi Shinzawa ◽  
Murat Dogru ◽  
Seika Den ◽  
Takehiro Ichijima ◽  
Kazunari Higa ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Dry Eye ◽  
Fatty Acid Binding Protein ◽  
Binding Protein ◽  
Sjögren Syndrome ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sjogren Syndrome ◽  
Fatty Acid Binding ◽  
Acid Binding Protein ◽  
Healthy Control

Purpose: Sjögren syndrome (SS) is a chronic inflammatory autoimmune disease of the lacrimal and salivary glands. This study compared the concentrations of epidermal fatty-acid binding protein (E-FABP) in the saliva, serum, and tears of SS patients with dry eye and dry mouth, with those of healthy adults to investigate the usefulness of E-FABP as a diagnostic marker for SS. Design: Prospective, observational case series. Participants: The subjects were 11 new patients with untreated Sjogren syndrome and 12 healthy control individuals. Methods: The diagnosis of SS was in accordance with the Ministry of Health, Labour and Welfare (Japan) Diagnostic Criteria (1999). Saliva, serum, and tear specimens were collected during internal medicine, dental, and ophthalmological examinations. The ophthalmological tests included the Dry Eye-related Quality of life Score (DEQS), tear break-up time (BUT), vital staining with fluorescein (FS) and lissamine green (LG), and the Schirmer test-1. The E-FABP concentration in the tears, saliva, and serum was measured by enzyme-linked immunosorbent assay (ELISA). Main outcome measure: The E-FABP concentrations were compared between patients and controls. Results: There were significant differences between the patient and healthy control groups in all ophthalmological test results. There were no significant differences between the groups in the E-FABP concentrations in the saliva (p = 0.1513) or the serum (p = 0.4799), but the E-FABP concentration in the tears significantly differed between groups. The E-FABP concentration in tears tended to be significantly lower in patients with SS (mean, 323.5 ± 325.6 pg/mL) than healthy control subjects (mean, 4076 pg/mL; p = 0.0136). The E-FABP concentration in tears significantly correlated with the results of dry eye parameters. Conclusion: The E-FABP concentration in tears appears to be related to ocular surface epithelial damage and tear stability and may be a promising novel biomarker in the diagnosis of SS.

Clinical utility of urinary liver-type fatty acid binding protein measured by latex-enhanced turbidimetric immunoassay in chronic kidney disease

2016 ◽  
Vol 54 (10) ◽  
Author(s):  
Atsuko Kamijo-Ikemori ◽  
Takeshi Sugaya ◽  
Maki Yoshida ◽  
Seiko Hoshino ◽  
Satoshi Akatsu ◽  
...  
Keyword(s):  
Chronic Kidney Disease ◽  
Fatty Acid ◽  
Kidney Disease ◽  
Clinical Utility ◽  
Fatty Acid Binding Protein ◽  
Binding Protein ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fatty Acid Binding ◽  
Acid Binding Protein

AbstractBackground:Urinary liver-type fatty acid binding protein (L-FABP) measured by enzyme-linked immunosorbent assay method (ELISA) was approved as a clinical biomarker of tubular damage by the Japanese Ministry of Health, Labor and Welfare (MHLW) in 2011. We evaluated a new latex-enhanced immunoturbidimetric assay (LTIA) to evaluate the clinical utility of urinary L-FABP measured by LTIA versus an ELISA assay.Methods:LTIA with anti-human L-FABP mouse monoclonal antibodies was performed using an automated clinical chemistry analyzer. Five positive samples with low, medium and high L-FABP concentrations were analyzed to determine the within-run precision. In patients with chronic kidney disease (CKD) (n=91), urinary L-FABP levels were measured by ELISA and LTIA.Results:Measurement of urinary L-FABP revealed urinary L-FABP levels within 30 min. The within-run coefficient of variation was 10.0% for 1.4 ng/mL, 4.4% for 2.5 ng/mL, 3.2% for 9.8 ng/mL, 1.5% for 50.1 ng/mL, and 1.2% for 102.7 ng/mL. Concentrations of urinary L-FABP measured by LTIA were significantly correlated with those measured by ELISA (ρ=0.932). Proportional systematic error was almost within limits of agreement (LOA). Urinary L-FABP levels measured by LTIA were significantly correlated with urinary albumin (ρ=0.634), urinary NAG (ρ=0.688) and eGFR (ρ=–0.561).Conclusions:Measurement of urinary L-FABP by LITA was simple, speedy, and similar in quality to ELISA results. Therefore, this method was approved as external body diagnosing medicines by the Japanese MHLW in 2014. Urinary L-FABP is expected to be widely used in various pathophysiological conditions by measuring urinary L-FABP using LTIA.

scholarly journals Evaluation of a sandwich enzyme-linked immunosorbent assay for the measurement of serum heart fatty acid-binding protein

2002 ◽  
Vol 39 (4) ◽  
pp. 404-405 ◽  
Author(s):  
Franca Pagani ◽  
Roberto Bonora ◽  
Graziella Bonetti ◽  
Mauro Panteghini
Keyword(s):  
Fatty Acid ◽  
Immunosorbent Assay ◽  
Fatty Acid Binding Protein ◽  
Binding Protein ◽  
Characteristic Curve ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fatty Acid Binding ◽  
Reference Limit ◽  
Acid Binding Protein ◽  
Upper Reference Limit

Background: We evaluated the sandwich enzyme-linked immunosorbent assay (ELISA) MARKIT®-M for the determination of heart fatty-acid-binding protein (H-FABP). Results and Conclusions: The between-run coefficient of variation of this assay was <3·9 and it showed good correlation with a previously established ELISA method. The upper reference limit in 30 healthy individuals was 6·1 μg/L. Admission serum H-FABP was evaluated against myoglobin in 41 patients with suspected myocardial infarction (onset of symptoms ≤ 5 h). H-FABP showed the same diagnostic efficiency as myoglobin [area (standard error) under the receiver operating characteristic curve: 0·798 (0·079) for H-FABP, 0·771 (0·085) for myoglobin, P = 0·55]. However, using the upper reference limit as decision cut-off, the sensitivity for H-FABP [91%; 95% confidence interval (CI): 76-98%] was significantly ( P = 0·019) higher than that of myoglobin (65%; 95% CI: 47-80%).

One-Step Enzyme-Linked Immunosorbent Assay (ELISA) for Plasma Fatty Acid-Binding Protein

1997 ◽  
Vol 34 (3) ◽  
pp. 263-268 ◽  
Author(s):  
K Will H Wodzig ◽  
Maurice M A L Pelsers ◽  
Ger J van der Vusse ◽  
Werner Roos ◽  
Jan F C Glatz
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Binding Protein ◽  
Rapid Determination ◽  
Binding Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fatty Acid Binding ◽  
Standard Curve ◽  
Acid Binding Protein ◽  
Sandwich Type ◽  
One Step

To allow a more rapid determination of heart-type fatty acid-binding protein (FABP) concentration in plasma a direct non-competitive (sandwich-type) ELISA was developed which uses high-affinity monoclonal antibodies to FABP. Total performance time of the one-step immunoassay is 45min. The standard curve was linear between 0·2–6μg/L, and the within-run and between-run coefficients of variations were below 6 and 11%, respectively. The serum FABP concentration measured in 79 healthy individuals was 1·6 (0·8) [mean (SD), range 0·3–5·0] μg/L. The assay can be used for rapid plasma or serum FABP measurement in the early diagnosis of acute myocardial infarction.

Preparation and characterization of a monoclonal antibody to the fatty acid binding protein (MFABP)

1989 ◽  
Vol 9 ◽  
pp. S25 ◽  
Author(s):  
H.E. Diede ◽  
M. Schrader ◽  
B. Zimmerbeutel ◽  
H. Hoppeler ◽  
M. Manns ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Fatty Acid ◽  
Fatty Acid Binding Protein ◽  
Binding Protein ◽  
Fatty Acid Binding ◽  
Acid Binding Protein
Sign in / Sign up

Export Citation Format

Share Document