Development of a Monoclonal Antibody-Based cELISA for the Analysis of Sulfadimethoxine. 2. Evaluation of Rapid Extraction Methods and Implications for the Analysis of Incurred Residues in Chicken Liver Tissue

2000 ◽  
Vol 48 (2) ◽  
pp. 545-550 ◽  
Author(s):  
Mark T. Muldoon ◽  
Sandra A. Buckley ◽  
Sudhir S. Deshpande ◽  
Carol K. Holtzapple ◽  
Ross C. Beier ◽  
...  
2020 ◽  
Vol 5 (2) ◽  
pp. 95 ◽  
Author(s):  
Rajashree Chowdhury ◽  
Prakash Ghosh ◽  
Md. Anik Ashfaq Khan ◽  
Faria Hossain ◽  
Khaledul Faisal ◽  
...  

To detect Post-kala-azar leishmaniasis (PKDL) cases, several molecular methods with promising diagnostic efficacy have been developed that involve complicated and expensive DNA extraction methods, thus limiting their application in resource-poor settings. As an alternative, we evaluated two rapid DNA extraction methods and determined their impact on the detection of the parasite DNA using our newly developed recombinase polymerase amplification (RPA) assay. Skin samples were collected from suspected PKDL cases following their diagnosis through national guidelines. The extracted DNA from three skin biopsy samples using three different extraction methods was subjected to RPA and qPCR. The qPCR and RPA assays exhibited highest sensitivities when reference DNA extraction method using Qiagen (Q) kit was followed. In contrast, the sensitivity of the RPA assay dropped to 76.7% and 63.3%, respectively, when the boil & spin (B&S) and SpeedXtract (SE) rapid extraction methods were performed. Despite this compromised sensitivity, the B&S-RPA technique yielded an excellent agreement with both Q-qPCR (k = 0.828) and Q-RPA (k = 0.831) techniques. As expected, the reference DNA extraction method was found to be superior in terms of diagnostic efficacy. Finally, to apply the rapid DNA extraction methods in resource-constrained settings, further methodological refinement is warranted to improve DNA yield and purity through rigorous experiments.


2012 ◽  
Vol 95 (2) ◽  
pp. 372-376 ◽  
Author(s):  
Elisabeth Halbmayr-Jech ◽  
Elisabeth Hammer ◽  
Richard Fielder ◽  
Jacqueline Coutts ◽  
Adrian Rogers ◽  
...  

Abstract In this work, a monoclonal antibody called G12, raised against the most immunotoxic peptide to celiac disease patients, was used to develop a sandwich ELISA. Preliminary results on cross-reactivities, recoveries, and extraction methods of the new assay are presented. The assay calibration was performed using material from the Prolamin Working Group. The antibody's specificity was determined by cross-reactivity studies on different grains, nuts, oils, and starches. Recovery of the assay was determined by spiking experiments on common food matrixes, as well as on problematic matrixes. Furthermore, sample extraction methods using ethanol, cocktail solution, and a proprietary buffer have been compared.


2011 ◽  
Vol 110 (2) ◽  
pp. 313-319 ◽  
Author(s):  
Hafeez-Ullah ◽  
M. Atif ◽  
S. Firdous ◽  
M. S. Mehmood ◽  
M. Y. Hamza ◽  
...  

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