The Sensitization Profile for Selected Food Allergens in Polish Children Assessed with the Use of a Precision Allergy Molecular Diagnostic Technique.

Individual populations show a variety of sensitization patterns, which may be associated with the geographic region, climate, dietary habits, or ways of preparing food. The purpose of this study was to comprehensively assess the food allergy sensitization profile in Polish children, particularly to eight food allergens (so-called "the Big 8"): cow milk, eggs, wheat, soybeans, fish, crustacean shellfish, tree nuts, and peanuts. To assess the prevalence and serum levels of specific immunoglobulins E (sIgE), we analyzed the results obtained from selected laboratories located in all regions of Poland that used the multiplex ALEX® test in the period from 2019 to 2022. Results from 3715 children were obtained. The mean age of the study population was 7.0 years. The results were stratified by age: <12 months (3.63%), 1-5 years (39.54%), 6-13 years (46.32%), and 14-18 years (10.0%). The final analysis included the sIgE results obtained with 95 food extracts and 77 food allergen molecules. The highest rates of sIgE to food allergen extracts were found for peanut (29.20%), hazel (28.20%), and apple (23.60%), and those to allergenic molecules were found for the PR-10 family of molecules (Cor a 1.0401 (23.77%), Mal d 1 (22.37%), Ara h 8 (16.93%), and globulin 7/8S (Ara h 1; 15.59%)). The lowest rates of sIgE reactivity to extracts were found for strawberry (0.40%), oregano (0.30%), and thornback ray (0.16%), and those to allergenic molecules were found for Mal d 2 (0.27%) (thaumatin-like protein, TLP), Ani s 1 (0.30%) (Kunitz-type serine protease inhibitor), and Che a 1 (0.43%) (Ole e 1 family). The rates of sensitization to storage proteins of the analyzed "the Big 8" molecules decreased significantly (p < 0.05) with age. Conversely, the rates of sensitization to PR-10 family proteins increased significantly with age. The three most common allergens in Poland, regardless of whether IgE was assayed against extracts or molecules of food allergens, were peanut, hazel, and apple (in different order depending on the ranking). A detailed analysis of sensitization to the extracts and molecules of main food allergens based on the results of a multiplex ALEX® test demonstrated the sensitization profile in Polish children (including molecular sensitization, particularly the "the Big 8" food allergen molecules), which shows considerable differences in comparison with those in other countries. Serum sIgE analysis of children from all regions of Poland revealed a food allergen molecular sensitization profile that changes with age.

The real life performance of 7 automated anti-SARS-CoV-2 IgG and IgM/IgA immunoassays.

OBJECTIVES: This study was aimed at providing some insights into the real-life performance of the commercial, clinically validated anti-SARS-CoV-2 antibody assays. METHODS: The residual, anonymized samples from 97 patients referred for anti-SARS-CoV-2 antibodies testing were included in the study. The initial assessment was performed with the Euroimmun ELISAs, followed by the assays provided by: NovaTec, Snibe, Vircell, Roche, Abbott and DiaSorin. The analyses of the results were performed separately for the antibodies of the early (IgM/IgA) and late (IgG) immune response. RESULTS: We observed a high variability of the results obtained with the investigated immunoassays. The fully concordant results were reported for only 57 out of 97 samples tested for IgG antibodies and for 34 out of 97 samples for IgM/IgA. The highest percentage of positive results was noted for the Euroimmun and Vircell ELISAs and the lowest for Novatec ELISAs.We proposed to distinguish true and false positive results based on the sum of positive results obtained with different methods. We arbitrarily considered reference positive samples reactive in at least half of the assays. The assay that proved to correlate the best with those reference results was the Roche electrochemiluminescence immunoassay. CONCLUSIONS: The differences observed between immunoassays targeting the early phase antibodies were much more pronounced than between IgG assays, suggesting their lower value for clinical use. Our study also showed a high percentage of plausibly false (positive or negative) results obtained with ELISAs, which suggests their inferiority to the automated immunoassays.

[Designing and optimization of real-time RT-PCR technique for the detection of hepatitis C virus (HCV) genome in blood serum as internal laboratory quality control]. / Opracowanie i optymalizacja techniki real-time rt-pcr do detekcji genomu wirusa zapalenia watroby typu c (HCV) w surowicy krwi dla celów kontroli wewnatrzlaboratoryjnej.

The need of hepatitis C virus (HCV) monitoring in serum samples of infected persons is of particular importance, because of chronic and non-symptomatic disease course of hepatitis C infection. We developed a novel "in-house" method variant for the detection of HCV genetic material in human blood serum. Detection technique is based on reverse transcription-real time polymerase chain reaction (RT-rPCR). We designed and analyzed several HCV 5' UTR-complementary PCR starter and probe sequence sets and we chose one set of highest HCV detection potency. Optimal concentration of starters and probe has been found. The 226-base pair long fragment of constitutively expressed glyceraldehyde 3-phosphate dehydrogenase gene served as internal endogenous control and should be added to each analysis in order to ensure that no PCR inhibitors are present. All parameters were optimized for Mx3005 QPCR System (Agilent Technology).