Utilization of a direct sandwich ELISA kit for rapid diagnosis of adenovirus hemorrhagic cystitis in a patient with adult T-cell leukemia
We report on a case of adenovirus hemorrhagi cystitis that developed in a 49-year-old woman during intensive chemotherapy for adult T-cell leukemia. Although rapid etiologic diagnosis is essential for the effective management of viral hemorrhagic cystitis, the isolation of adenovirus from urine is often too time-cousuming. We detected the adenovirus antigen in the patient’s urine using an Adenoclone direct sandwich ELISA kit (Cambridge Bio Science), which is commonly employed for the diagnosis of adenovirus conjunctivitis. Treatment consisted of intravenous vidarabine and bladder irrigation, which resulted in prompt clinical improvement. The Adenoclone kit was useful for the rapid etiologic diagnosis of adenovirus hemmorrhagic cystitis.
Determination of okadaic acid content of dinoflagellate cells: a comparison of the HPLC-fluorescent method and two monoclonal antibody ELISA test kits.
Total okadaic acid (okadaic acid plus methylokadaic acid) in acclimated clones of the dinoflagellates Prorocentrum hoffmannianum and P. lima was determined using the HPLC-fluorescent method, UBE ELISA test kit, and Rougier ELISA test kit. The nonokadaic acid-producing species. Amphidinium klebsii, Prorocentrum mexicanum, P. micans, P. cassubicum, and Gambierdiscus toxicus were examined using the same methods of analysis. All three methods yielded consistent results for P. hoffmannianum which produces only okadaic acid. However, results of the three methods were not consistent for P. lima which produces both okadaic acid and methylokadaic acid. The UBE ELISA demonstrated little or no cross-reactivity with methylokadaic acid; whereas, the Rougier ELISA demonstrated varying degrees of cross-reactivity with that analog. Analyses of nonokadaic acid producing-species yielded negative results, with one exception. The Rougier ELISA demonstrated reactivity with extracts of G. toxicus. Since outbreaks of DSP may be caused by okadaic acid, methylokadaic acid, or a combination of these toxins, both ELISA kits may underestimate total toxin present in toxic shellfish.
Simplified validation of the ELISA kit determination of Microcystins in surface water
The enzyme-linked immunosorbent assay (ELISA), as a universal method for the determination of Microcystins, is of great significance for the rapid detection of Microcystins pollution. This study aimed to propose a simplified validation method for Microcystins ELISA kit by summarizing related documents and guidelines. After summarizing and clarifying from 20 validation parameters, 11 parameters were selected to simplify the validation of Microcystins ELISA kit. In addition, the acceptable range and validation details of each parameter were analyzed. The results indicated that the coefficient of determination of the Microcystin-LR standard curve was higher than 0.99. The concentration of quality control samples was within control limits.
The accuracy of spiked and proficient samples was within 70%-130%. The variability of intra-assay, inter-assay, and reproducibility was less than 11, 15 and 21%, respectively. The LOD and LLOQ were 0.002 μg/L and 0.05 μg/L, respectively. When the concentration of Microcystins exceeded 5 μg/L, it was recommended to dilute the samples to the working range before detection. The specificity was estimated with seven Microcystin analogues and three amino acids, indicating that the cross-reactivity was less than 30%. These results revealed that the ELISA kit was satisfactory for detecting Microcystins in water.
Can Procalcitonin Be Dosed in Bovine Milk Using a Commercial ELISA Kit?
The aim was to evaluate the use of a bovine procalcitonin (PCT) ELISA kit (Cusabio, China) for assessing PCT in bovine milk samples. Validation was performed by using 10 plasma and corresponding milk samples from mastitic cows. The limit of detection (LOD) was calculated. The coefficient of variation (CV%) of the readings of five plasma samples measured five times in the same plate (intra-assay) and the CV% of the same five samples read five times in three separate plates was evaluated. Parallelism was determined by serial twofold dilutions of five plasma and corresponding milk samples.
Milk samples were analyzed with and without centrifugation. Regarding plasma PCT, the method presented an inter- and intra-CV < 23.7% and parallelism had very good recovery values. The ELISA kit studied can measure bovine plasma PCT concentrations. The kit antibodies fail in binding PCT in milk samples because all centrifuged milk samples showed a lower LOD than blank samples. Only three uncentrifuged milk samples showed measurable PCT concentrations. Due to these results, the commercial ELISA kit investigated could not be employed for the detection of PCT in milk samples.
Quantification of Barley Contaminants in Gluten-Free Oats by Four Gluten ELISA Kits
Pure oats are generally accepted to be safe for most celiac patients, and consumption of oats provides advantageous dietary fibers. However, oats can be contaminated by gluten proteins from wheat, barley, and/or rye. The analytical challenge lies in the reliability of the quantification method and how to maintain the contamination level under a gluten-free food threshold of 20 mg/kg. In this study, we investigated barley-spiked oat flour samples at four levels using four gluten ELISA kits. The largest recovery variance was with the R5 kit that gave 5-6 times overestimation; the G12 kit cross-reacted with oat proteins and gave 4-5 times overestimation at all spiked levels. The Total Gluten and Morinaga kits gave satisfactory recoveries.
Total barley hordeins were isolated and characterized to be used as a common calibrator in all four kits aiming at harmonizing the results and to test the kits’ performance. Immunoblotting of total hordein isolate revealed that Total Gluten and Morinaga antibodies provided an overall detection, while R5 and G12 antibodies recognized specific hordein groups leading to a larger difference when wheat and barley were used as the calibrant. Calibration with total hordein isolate corrected the overestimation problem and decreased the variability between the four gluten kits.
Evaluation of a genus-specific rGroEL 1-524 IgM-ELISA and commercial ELISA kits during the course of leptospirosis in Thailand
In the present study, we developed a genus-specific rGroEL1-524 IgM-ELISA assay for use in screening diagnosis of suspected leptospirosis among acute undifferentiated febrile illness patients during acute fever. The diagnostic accuracies of the rGroEL1-524 IgM-ELISA, commercial Panbio IgM-ELISA, and Virion-Serion Classic IgG-ELISA were evaluated using 133 Thai leptospirosis sera and 210 controls. Sensitivities were 91.7%, 59.6%, and 17.7% for acute infection, and the specificities were 92.6%, 90.2%, and 88.3% for the non-leptospirosis control, respectively. The rGroEL1-524 IgM-ELISA had high sensitivity, at 92.3% and 91.7%, among culture-positive and MAT-negative cases at 1-3 days post-onset of symptoms (DPO1-3), respectively.
Vero Cell HCP ELISA kit | ||||
| F500 | Cygnus Technologies | 1 kit (96 wells plate) | 1296 EUR | |
Anti-Vero Cell HCP | ||||
| VC807 | Cygnus Technologies | 1 ml | 400.8 EUR | |
Anti-Vero Cell HCP | ||||
| VC807-AF | Cygnus Technologies | 1 mg | 2295.6 EUR | |
Anti-Vero Cell HCP | ||||
| VC807AF-HRP | Cygnus Technologies | 1 mg | 4882.8 EUR | |
VERO Host Cell Proteins (HCPs) ELISA Kit, 96 tests, Quantitative | ||||
| 770-165-HCP | Alpha Diagnostics | 1 kit | 927.6 EUR | |
anti-Vero Cell HCP, Affinity Purified | ||||
| VC807-AFB | Cygnus Technologies | 1 mg | 4143.6 EUR | |
Anti-Vero Cell HCP, IgG, Goat | ||||
| VC807-PA | Cygnus Technologies | 1 ml | 585.6 EUR | |
Anti-Vero Cell HCP affinity purified | ||||
| VC807.5-AF | Cygnus Technologies | 0.5 mg | 1926 EUR | |
Vero Cell HCP Standards Set, A-F | ||||
| F503 | Cygnus Technologies | 1 ml | 540 EUR | |
Saccharomyces Cerevisiae Host Cell Proteins (SC-HCP) | ||||
| 760-160-HCP | Alpha Diagnostics | 500 ug | 1016.4 EUR | |
CAPÐ’ cell line HCP ELISA Kit | ||||
| F820 | Cygnus Technologies | 1 kit (96 wells plate) | 1296 EUR | |
Vero cell Standard F | ||||
| F503F | Cygnus Technologies | 1 ml | 234 EUR | |
Vero Cell Antigen Concentrate | ||||
| F503H | Cygnus Technologies | 400 ul | 678 EUR | |
Vero Cell Control Antigen | ||||
| F507 | Cygnus Technologies | 60 ul | 678 EUR | |
CHO HCP ELISA kit | ||||
| F015 | Cygnus Technologies | 1 kit (96 wells plate) | 1296 EUR | |
S.cerevisiae HCP ELISA kit | ||||
| F135 | Cygnus Technologies | 1 kit (96 wells plate) | 1296 EUR | |
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Impaired specificity on scrub typhus was found, possibly from antibody cross-reaction to ortholog GroEL. Commercial Panbio IgM-ELISA had sensitivities at DPO1-3 of 30.8% and 41.7% for culture-positive and MAT-negative cases whereas Virion-Serion IgG-ELISA showed sensitivities of 5.9% and 13.3%, respectively. The rGroEL1-524 IgM-ELISA could be useful as a screening test for early diagnosis. The performance of the commercial ELISA suggests the applicability of IgM-ELISA for diagnosis, while IgG-ELISA is useful for seroprevalence surveys. However, confirmation by reference tests is recommended.