Gluten Assessment in Beers: Comparison by Different Commercial ELISA Kits and Evaluation of NIR Analysis as a Complementary Technique

Traditionally, beers are made with gluten-containing cereals. It is crucial to have rapid analytical methodologies that allow gluten content control of the beers for celiac consumers. We assess the content of gluten in 65 conventional and 41 gluten-free labeled beers commercialized in Europe and compare the results in a subgroup of 71 beers with three ELISA kits. This research allows gathering information on the potential complementary utility of NIR analysis applied to gluten analysis of gluten-free beers in terms of time saving.
Results obtained with the ELISA technique identified competitive R5 to be the most sensitive in detecting the prolamins, by eliciting a higher number of beers containing gluten above 20 mg/kg. The gluten content in conventional beers tested increased with the presence of wheat as raw material and with the use of ale-type yeasts. By using competitive R5, 3 out of the 41 gluten-free labeled beers appeared to contain gluten above 20 mg/kg, and conversely, 15 out of 65 of the conventional beers showed a gluten content below this threshold. According to our approaches, NIR did not achieve a suitable correlation with ELISA results, neither for gluten quantification nor for discrimination, and therefore, it cannot be proposed as a complementary technique.

Evaluation of Loopamp™ Leishmania Detection Kit and Leishmania Antigen ELISA for Post-Elimination Detection and Management of Visceral Leishmaniasis in Bangladesh

  • With reduced prevalence of visceral leishmaniasis (VL) in the Indian subcontinent (ISC), direct and field deployable diagnostic tests are needed to implement an effective diagnostic and surveillance algorithm for post-elimination VL control. In this regard, here we investigated the diagnostic efficacies of a loop-mediated isothermal amplification (LAMP) assay (Loopamp™ Leishmania Detection Kit, Eiken Chemical CO., Ltd, Japan), a real-time quantitative PCR assay (qPCR) and the Leishmania antigen ELISA (CLIN-TECH, UK) with different sampling techniques and evaluated their prospect to incorporate into post-elimination VL control strategies. Eighty clinically and rK39 rapid diagnostic test confirmed VL cases and 80 endemic healthy controls were enrolled in the study.
  • Peripheral blood and dried blood spots (DBS) were collected from all the participants at the time of diagnosis. DNA was extracted from whole blood (WB) and DBS via silica columns (QIAGEN) and boil & spin (B&S) methods and tested with qPCR and Loopamp. Urine was collected from all participants at the time of diagnosis and was directly subjected to the Leishmania antigen ELISA. 41 patients were followed up and urine samples were collected at day 30 and day 180 after treatment and ELISA was performed. The sensitivities of the Loopamp-WB(B&S) and Loopamp-WB(QIA) were 96.2% (95% CI 89·43-99·22) and 95% (95% CI 87·69-98·62) respectively. The sensitivity of Loopamp-DBS(QIA) was 85% (95% CI 75·26- 92·00).
  • The sensitivities of the qPCR-WB(QIA) and qPCR-DBS(QIA) were 93.8% (95% CI 86·01-97·94) and 72.5% (95% CI 61·38-81·90) respectively. The specificity of all molecular assays was 100%. The sensitivity and specificity of the Leishmania antigen ELISA were 97.5% (95% CI 91·47-99·70) and 91.95% (95% CI 84·12-96·70) respectively. The Leishmania antigen ELISA depicted clinical cure at day 180 in all the followed-up cases. Efficacy and sustainability identify the Loopamp-WB(B&S) and the Leishmania antigen ELISA as promising and minimally invasive VL diagnostic tools to support VL diagnostic and surveillance activities respectively in the post-elimination era.

Determination of Aflatoxin M1 in Raw Milk Using an HPLC-FL Method in Comparison with Commercial ELISA Kits-Application in Raw Milk Samples from Various Regions of Greece

The highly toxic Aflatoxin M1 (AFM1) is most often detected in milk using an Enzyme-Linked-Immunosorbent Assay (ELISA) for screening purposes, while High-Performance Liquid Chromatography with Fluorescence Detector (HPLC-FL) is the reference method used for confirmation. The aim of the present study was the comparison between three commercially available ELISA kits and a newly developed HPLC-FL method for the determination of the AFM1 in milk samples. The developed HPLC-FL method was validated for the AFM1 and Aflatoxin M2 (AFM2), determining the accuracy, precision, linearity, decision limit, and detection capability with fairly good results.
All three ELISA kits were also validated and showed equally good performance with high recovery rates. Moreover, the Limit Of Detection (LOD) and Limit Of Quantification (LOQ) values were found to be significantly lower than the Maximum Residue Limit (MRL) (50 ng kg-1). After the evaluation of all three commercial kits, the ELISA kit with the optimum performance along with the HPLC method was used for the determination of AFM1 in raw cow’s, goat’s, and sheep’s milk samples (396) obtained from producers in different regions of Greece. The evaluation of both methods showed that this ELISA kit could be considered as a faster and equally reliable alternative method to HPLC in routine analysis for the determination of AFM1 in milk.

T-Pro Plasmid Mini Kit (100)

RB94-YPD100 T-Pro Biotechnology 100preps/Kit 193.2 EUR

T-Pro Plasmid Mini Kit (250)

RB94-YPD250 T-Pro Biotechnology 250preps/Kit 266.4 EUR

T-Pro Plasmid Midi Kit (20)

RB94-YPI020 T-Pro Biotechnology 20preps/Kit 255.6 EUR

T-Pro Plasmid Maxi Kit (10)

RB94-YPM010 T-Pro Biotechnology 10preps/Kit 266.4 EUR

T-Pro BCA Protein Assay kit

JB04-D001 T-Pro Biotechnology 500assay/KIT 244.8 EUR

T-Pro Cell Counting Kit (CCK8)

JK95-C008L T-Pro Biotechnology 5ml*20/BT 1000 EUR

T-Pro Cell Counting Kit (CCK8)

JK95-C008M T-Pro Biotechnology 5ml*2/BT 120 EUR

T-Pro Cell Counting Kit (CCK8)

JK95-C008S T-Pro Biotechnology 1ml*5/BT 80 EUR

T-Pro LumiDura Chemiluminescence Detection Kit (ECL Kit)

JT96-K006M T-Pro Biotechnology 250ml*2/Kit 200 EUR

T-Pro LumiDura Chemiluminescence Detection Kit (ECL Kit)

JT96-K006S T-Pro Biotechnology 100ml*2/Kit 100 EUR

T-Pro Genomic DNA Mini Kit (100)

RB94-NGS100 T-Pro Biotechnology 100preps/kit 255.6 EUR

T-Pro Genomic DNA Midi Kit (20)

RB94-NGM020 T-Pro Biotechnology 20preps/kit 266.4 EUR

T-Pro Bradford Protein Assay kit(1X)

JB04-D002 T-Pro Biotechnology 500ml/BT 193.2 EUR

T-Pro Bradford Protein Assay kit (5X)

JB04-D003 T-Pro Biotechnology 500ml/BT 120 EUR

T-Pro LumiFast Plus Chemiluminescence Detection Kit (ECL Kit)

JT96-K002M T-Pro Biotechnology 250ml*2/Kit 244.8 EUR

T-Pro LumiFast Plus Chemiluminescence Detection Kit (ECL Kit)

JT96-K002S T-Pro Biotechnology 100ml*2/Kit 182.4 EUR

T-Pro LumiLong Plus Chemiluminescence Detection Kit (ECL Kit)

JT96-K004M T-Pro Biotechnology 250ml*2/Kit 286.8 EUR

T-Pro LumiLong Plus Chemiluminescence Detection Kit (ECL Kit)

JT96-K004S T-Pro Biotechnology 100ml*2/Kit 204 EUR

T-Pro Plant Genomic DNA Mini Kit (100)

RB94-PGS100 T-Pro Biotechnology 100preps/kit 266.4 EUR

T-Pro Gradient Gel Solution kit 4-12%

JB02-B110M T-Pro Biotechnology 250ml+100ml/kit 60 EUR

T-Pro Plant Genomic DNA Midi Kit (20)

RB94-PGM020 T-Pro Biotechnology 20preps/kit 266.4 EUR

T-Pro Endotoxin Removal Plasmid Midi kit (25)

RB94-EPI020 T-Pro Biotechnology 20preps/Kit 297.6 EUR

T-Pro Endotoxin Removal Plasmid Maxi kit (10)

RB94-EPM010 T-Pro Biotechnology 10preps/Kit 318 EUR

T-Pro Gel/PCR DNA Purification Mini Kit (100)

RB94-NES100 T-Pro Biotechnology 100preps/Kit 193.2 EUR

Comparison of a new rapid method for determination of serum anti-adalimumab and anti-infliximab antibodies with two established ELISA kits

Background: Adalimumab (ADL), infliximab (IFX) and their biosimilars are widely used biological drugs. Some patients, however, generate neutralizing antibodies that hamper the effectiveness of these drugs. Evidence shows therapeutic drug monitoring of serum levels ADL/IFX and anti-drug antibodies (ADA) is useful to improve treatment effectiveness. We evaluated a new rapid quantitative method, Quantum Blue (QB), for determining serum anti-ADL and anti-IFX antibodies (Research Use Only labelling) by comparing it with two established ELISA kits, Promonitor (PM) and Lisa-Tracker (LT).
Methods: Eighty samples (40 for each drug type) were analysed. Percentage of agreement and kappa statistic were used to compare positive/negative ADA results. Clinical implications for drug treatment in the patients with discordant results were evaluated. The Chi-square test was used to analyze differences for ADA detection in patients with disease flare and without concomitant immunosuppressant treatment.
Results: Agreement exceeded 80 % among anti-ADL methods. Although LT ELISA showed a lower capacity in detecting anti-ADL antibodies, discrepancies were found for levels close to the cut-off concentration, thus having minimal impact on clinical decisions. Conversely, QB anti-IFX displayed low agreement with PM and LT ELISA kits (67.5 % and 50 %, respectively), and was unable to detect high levels of antibodies, therefore having major clinical implications. Agreement between PM and LT ELISA anti-IFX kits was 82.5 % with all discordant results being undetected for PM and slightly positive for LT.
Conclusion: QB anti-ADL shows similar performance to ELISA kits while QB anti-IFX needs further improvements to achieve reliable antibody detection.

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