Repeatability of a Commercially Available ELISA Test for Determining the Herd-Level Salmonella enterica subsp. enterica Serovar Dublin Status in Dairy Herds Using Bulk Milk

August 23, 2021
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An Enzyme-Linked Immunosorbent Assay (ELISA) is at the moment accessible for detection of antibodies in opposition to Salmonella Dublin in bovine milk. Nonetheless, when utilized in a surveillance program, samples could endure numerous storage situations. The target of this examine was to estimate the repeatability of an ELISA take a look at when used on recent and frozen samples.
Every of 845 bulk milk collected samples was subdivided into Three aliquots and analyzed utilizing PrioCHECK™ Salmonella Ab Bovine Dublin. ELISA p.c positivity outcomes (PP%) had been in contrast between aliquots submitted to the preliminary evaluation and a second evaluation carried out 24 h later. The third aliquots had been both preserved for 13-14 days (n = 413) or 25-28 days (n = 432) at -20°C previous to evaluation and outcomes had been in comparison with the preliminary evaluation.
There was glorious concordance between the 2 preliminary values and with values obtained after 13-14 and 25-28 days-freezing. The corresponding concordance correlation coefficients had been 0.96, 0.97, and 0.94, respectively. Bland-Altman plots confirmed variations of PP% of 0.1 share factors on common between the preliminary and second recent samples. Freezing for 13-14 and 25-28 days led to overestimation of the preliminary values by 0.1, and 0.
Four share factors, respectively. Relating to the classification of samples, higher disagreement was noticed between 25 and 28 days-frozen and preliminary samples when utilizing the cut-off 15% (kappa = 0.76) in comparison with 35% (kappa = 0.90). Our examine confirmed that PrioCHECK™ has good repeatability and that frozen bulk milk samples might generate dependable outcomes. Nonetheless, the bigger variability at decrease PP% ought to be thought-about when organising a threshold.
Particular person and herd-level milk ELISA take a look at standing for Johne’s illness in Eire after correcting for non-disease-associated variables
Antibody-detecting exams for Mycobacterium avium ssp. paratuberculosis (MAP) have low sensitivity and imperfect specificity for detection of an infection. Sensitivity will increase because the illness progresses. Except for an infection standing and stage of illness, a number of elements have an effect on take a look at efficiency. These elements haven’t but been studied in dairy cows producing decrease volumes of milk with greater solids focus, corresponding to these managed in low-input, pasture-based manufacturing methods.
Moreover, the impact of correcting for these associations on particular person and herd take a look at standing can also be unknown. The primary goal of this examine was to look at the connection between MAP antibody response in milk and milk yield, somatic cell rely (SCC), fats and protein contents, and stage of lactation in dairy cows enrolled within the nationwide Johne’s Illness Management Programme (JDCP) in Eire.
The second goal was to look at the impact of correcting the antibody response for these associations on the take a look at standing of particular person cows and herds, provided that particular person exams are sometimes used to outline a herd’s standing. Information had been extracted for herds within the JDCP from January 2014 to December 2015 inclusive, consisting of 42,657 milk recordings from 18,569 cows throughout 187 dairy herds.
Two linear regression fashions had been constructed to analyze the affiliation between log-transformed MAP sample-to-positive ratio and milk recording knowledge and in primi- and multiparous cows. Days in milk was modeled as a B-spline in every mannequin, and cow and herd had been included as random results. Throughout each fashions, pure log-transformed MAP antibody response was negatively related to milk yield, positively related to protein and fats manufacturing, and had a curvilinear affiliation with log-transformed SCC.
The affiliation between MAP antibody response and days in milk diversified over the course of the lactation. Nonetheless, when mixed, these variables defined solely 5.1% of the variation within the antibody response of the inhabitants. After correcting for these associations, 93 multiparous cows and 20 primiparous cows modified class (unfavorable, suspect, or optimistic).

When thought-about on the herd-test stage, out of a complete of 531 herd exams, 1 herd modified from unfavorable to optimistic, and 5 herds modified from optimistic to unfavorable. This examine gives helpful data to help within the interpretation of antibody outcomes for herds testing animals for the presence of MAP an infection. At an total inhabitants stage, correction of the serological response for non-disease-associated elements has the potential to vary the standing of solely a small variety of cows.
On the herd stage, the proportion of herds altering standing was minimal. Nonetheless, relying on the implications of a herd-level serological prognosis, consideration ought to be given to correcting for these non-disease-associated variables inside the context of nationwide JD management packages.
DIAGNOSTIC PARAMETERS OF IN VIVO (SKIN PRICK) AND IN VITRO (ELISA) TESTS FOR DETERMINATION OF EPIDERMAL CAT AND DOG ALLERGENS SENSITIZATION IN PATIENTS WITH ALLERGIC RHINITIS AND ATOPIC ASTHMA
Goal was to review and evaluate the parameters of the specificity and sensitivity of pores and skin testing and serologic willpower of particular cat and canine IgE. 88 sufferers with allergic rhinitis and / or bronchial asthma had been examined by three totally different strategies of particular allergic prognosis (in vivo and in vitro) in accordance with the rules of the ethics committee of the Nationwide Pirogov memorial medical college, all had been past the acute interval. The inclusion standards had been allergic rhinitis prognosis (each intermittent and protracted) and \ or bronchial asthma. Pores and skin prick take a look at was carried out based on the classical testing process in accordance with regulatory paperwork with business extracts of allergens.
Western blot testing for particular IgE ranges was carried out utilizing RIDA qLine take a look at methods (R-Biopharm AG, Darmstadt, Germany) and Euroline (Euroimmun). The sIgE focus was transformed to a nominal scale (grades) based on the next guidelines: < 0.35 IU mL-1-level 0 (unfavorable), (0.36-0.69) IU mL-1-level 1 (boundary ranges ), (0.7- 3.49) IU mL-1-level 2 (barely elevated), (3.50-17.4) IU mL-1-level 3 (reasonably elevated), (17.5-49 , 9) IU mL-1-level 4 (excessive ranges), (50-100) IU mL-1-level 5 (very excessive ranges) and > 100 IU mL-1-level 6 (extraordinarily excessive ranges).
Thus, the outcomes of the 2 methods for the willpower of particular IgE for canine allergen by the Rida AllergyScreen and Euroline strategies don’t agree very properly as a result of systematic divergence of indicators; the outcomes of the 2 methods for the willpower of particular IgE for cat allergen by the Rida AllergyScreen and Euroline strategies agree very properly.
There’s glorious settlement between the pores and skin take a look at with cat allergen and the detection of particular IgE by the Rida AllergyScreen take a look at, between the pores and skin take a look at with cat allergen and the detection of particular IgE by the Euroline methodology. There’s good settlement between the pores and skin take a look at with canine wool allergens and the detection of particular IgE by the Rida AllergyScreen take a look at, between the pores and skin take a look at with canine hair allergen and the detection of particular IgE by the Euroline methodology there’s passable settlement.
BCA Protein Assay Reagent A |
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K544-1000 | Biovision | each | EUR 901.2 |
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PCR0352 | Scientific Laboratory Supplies | PK100 | EUR 454.8 |
AlamarBlue Cell Viability Assay Reagent |
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AR4002-10ml | BosterBio | 10ml | EUR 184.8 |
AlamarBlue Cell Viability Assay Reagent |
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AR4002-25ml | BosterBio | 25ml | EUR 338.4 |
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1996-2 | Biovision | each | EUR 124.8 |
T-Pro BCA Protein Assay Reagent A |
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T-Pro BCA Protein Assay Reagent B |
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P8101-050 | GenDepot | 450ml | EUR 225.6 |
human granzyme B activity assay reagent set |
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QZBGrB2 | Quickzyme | 2 plates 192 assays | EUR 660 |
Polystyrene-bottomed 96-well plate (ELISA) |
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CT361 | U-CyTech | 6 | EUR 87.6 |
Bicinchoninic Acid(BCA) Protein Assay Reagent B |
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P8102-001 | GenDepot | 10ml | EUR 165.6 |
6-well reagent vessel for IsoPure Mini, cs 100 |
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CytoSelect Cell Proliferation Assay Reagent (Fluorometric) |
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CBA-250 | Cell Biolabs | 10 mL | EUR 490.8 |
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96-Well Filter Plate |
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Description: Suitable for cell culture but only for suspension cells
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96 well filter plate (960ul each well) |
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Description: PCR Plates & Tubes; A354 PCR Plate-Axygen |
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PFS096 | ProFoldin | kit | EUR 347.84 |
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1011601 | Scientific Laboratory Supplies | 1L | EUR 52.8 |
Human Topoisomerase II DNA Decatenation Assay Reagent Set (enzyme not included) |
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5 x Clear 96 well Plate, 5EA |
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X018-2EA | Arbor Assays | 2EA | EUR 13 |
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X018-4EA | Arbor Assays | 4EA | EUR 31 |
Protein A-coated ELISA plate (8 well strips, 96 wells/plate) 5 plates/pack |
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Protein G-coated ELISA plate (8 well strips, 96 wells/plate) 5 plates/pack |
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96 well filter plate (no filter) |
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1056700 | Scientific Laboratory Supplies | 100ML | EUR 52.8 |
96 well DNA plate with membrane (960ul each well) |
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SD5007 | Bio Basic | 12UNIT | EUR 291.24 |
96 well RNA plate with membrane (960ul each well) |
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SD5009 | Bio Basic | 12UNIT | EUR 313.16 |
Protein A&G-coated ELISA plate (8 well strips, 96 wells/plate) 5 plates/pack |
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PRTAG75-5P | Alpha Diagnostics | 1 PK | EUR 343.2 |
Block-Free ELISA Reagent |
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M4102-250 | Biovision | each | EUR 542.4 |
Block-Free ELISA Reagent |
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M4102-50 | Biovision | each | EUR 176.4 |
96-Well Plate Viral DNA Miniprep Kit |
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VT92032 | Bio Basic | 2XPlates, 192prep | EUR 495.86 |
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1053601 | Scientific Laboratory Supplies | 100ML | EUR 435.6 |
96-Well Plate Plasmid DNA Miniprep Kit |
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BS415 | Bio Basic | 5XPlates, 480prep | EUR 300.37 |
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Benchmark Beadblaster 96 Eva Cap Mat for 96 Well Deep Well Plate |
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BS369 | Bio Basic | 5XPlates, 480prep | EUR 300.37 |
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EP Reagent Sulfomolybdic Reagent R3 |
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1086500 | Scientific Laboratory Supplies | 1L | EUR 354 |
EP Reagent Phosphomolybdotungstic Reagent |
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1065000 | Scientific Laboratory Supplies | 100ML | EUR 201.6 |
The BCS Screen HT-96 Single Reagent 100mL |
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M-MDSR-105 | MiTeGen | 100 ml ml | EUR 155 |
Description: The BCS Screen HT-96 Single Reagent 100mL |
96 WELL PLATE; SQUARE; U BASE; 1.6ML; PP; 96/PK |
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60180-P155 | Scientific Laboratory Supplies | PK96 | EUR 561.6 |
PCR Plate 96 Well No Skirt Nat |
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10 x ELISA Reagent Diluent |
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GR103028 | Genorise Scientific | 100 mL | EUR 77 |
96 WELL PLATE; SQUARE; 2ML; PP; 1/PK |
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60180-P403 | Scientific Laboratory Supplies | EACH | EUR 19.2 |
PVDF membrane-bottomed 96-well plate + lid (ELISPOT) |
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MULTIWELL PLATE POLYSTYRENE 96-WELL FLA |
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M2936-100EA | Scientific Laboratory Supplies | PK100 | EUR 131.87 |
96 WELL CLEAR PCR PLATE FOR ABI |
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PCR-96-AB-C | CORNING | 10/pk | EUR 200.4 |
Description: PCR Plates & Tubes; A354 PCR Plate-Axygen |
96-Well Plate PCR Products Purification Kit |
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BS365 | Bio Basic | 5XPlates, 480prep | EUR 300.37 |
96-Well Plate PCR Products Purification Kit |
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BS3652 | Bio Basic | 2xPlates, 192prep | EUR 186.19 |
96 Well ExoELISA plate (12x8 well strips, pack of 10 plates) |
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EXOEL-96P-1 | SBI | 10 Plates | EUR 255.6 |
1.2ML 96 WELL DEEP WELL PLATE HIGH CLARITY PRE-STERILIZED |
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P-DW-12-HC-S | CORNING | 5/pk | EUR 248.4 |
Description: Deep Well/Assay Plates - Axygen; Deep-well plates - Axygen |
96 Well PCR Plate Divisible In 8 |
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PCR1060 | Scientific Laboratory Supplies | PK50 | EUR 174 |
96 Well PCR Plate Divisible In 8 |
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PCR1062 | Scientific Laboratory Supplies | PK50 | EUR 187.2 |
The systematic error of the measurement outcomes between two in vitro exams for cat allergen was 0.1 ku/l, which signifies the presence of a small systematic distinction, the systematic error of the measurement outcomes between two in vitro exams for canine allergen was 0,26 ku/l, which signifies the presence of a reasonable systematic distinction.