The importance of EN ISO 15189 accreditation of allergen-specific IgE determination for reliable in vitro allergy diagnosis.

BACKGROUND: Allergen-specific serum immunoglobulin E detection and quantification have become an important step in allergy diagnosis and follow-up. In line with the current trend of laboratory test accreditation to international standards, we set out to design and assess an accreditation procedure for allergen-specific serum IgE. METHODS: Method validation according to the accreditation procedure under the EN ISO 15189 standard was carried out for allergen-specific immunoglobulin E determination using the fluoroimmunoenzymatic method ImmunoCAP(®) (ThermoFisher). Data were produced by 25 hospital laboratories in France. A total of 29 allergen specificities including mixes, extracts, and molecular allergens were assayed. Allergen-specific serum immunoglobulin E concentrations ranged from 0.1 to 100 kUA /l. RESULTS: Repeatability, reproducibility, and accuracy results fulfilled method validation criteria for automated laboratory tests and proved similar irrespective of the allergen specificity, allergen-specific serum immunoglobulin E concentration, or individual laboratory. CONCLUSION: Allergen-specific serum immunoglobulin E determination with the fluoroimmunoenzymatic method ImmunoCAP(®) is a highly repeatable, reproducible, and accurate method which may be considered as a single analyte assay in view of the EN ISO 15189 accreditation procedure.

Similar lipid profile but improved long-term outcomes with sirolimus after cyclosporine withdrawal compared to sirolimus with continuous cyclosporine.

Renal transplant recipients show an increased risk of cardiovascular disease compared with a nontransplant population. Herein we have shown an analysis of a randomized controlled trial wherein 525 patients receiving a first or second (9.7%) renal allograft from a deceased (89.1%), a living-related (7.8%), or a living-unrelated donor (3.1%) received sirolimus (SRL), cyclosporine (CsA), and steroids (ST) at the time of transplantation with randomization at 3 months after transplantation of 430 eligible patients to continue on SRL-CsA-ST or to have CsA withdrawn with increased SRL trough targets (SRL-ST group). Graft survival, patient survival, and renal function at 5 years were analyzed by average fasting total cholesterol (200 mg/dL) and triglyceride (240 mg/dL) subgroups. At 5 years, total, high-density lipoprotein (HDL), and low-density lipoprotein [LDL] cholesterol and triglyceride values were similar between the groups. Statins ( approximately 80% of patients of both groups) were most effective to lower cholesterol ( approximately 50 mg/dL; P < .001; both groups), and fibrates ( approximately 25% of patients of both groups) were most effective to decrease triglycerides ( approximately 100 mg/dL; P < .001; both groups). Renal function and blood pressure were significantly better with SRL-ST. Hypercholesterolemia and hypertriglyceridemia were associated with reduced graft survival, patient survival, and calculated GFR, but the only significant difference was lower graft survival among SRL-CsA-ST patients with hypertriglyceridemia. Cardiovascular-related deaths were reported in 3.7% and 2.8% of patients in the SRL-CsA-ST and SRL-ST groups, respectively. In conclusion, when compared with continuous SRL-CsA-ST, CsA withdrawal at 3 months followed by SRL-ST significantly improved glomerular filtration rate (GFR) and blood pressure without a further increase in lipid parameters or an incidence of untoward effects from hyperlipidemia, despite a 2-fold higher SRL exposure.

Immunochemical comparison of the allergenic potency of spores and mycelium of Cladosporium cladosporioides extracts by a nitrocellulose electroblotting technique.

BACKGROUND: The lack of well standardized or characterized extracts that contain the relevant allergens of the appropriate fungus is resulting in a high heterogeneity of the commercial preparation. MATERIAL AND METHODS: Immunochemical detection of the allergens composition of spore and mycelium of C. cladosporioides was studied by electroblotting using sera from Cladosporium allergic patients and 125 I- anti- human IgE. A MW range of allergens between 16 to 88 KDa was identified. The most important with a MW of 16, 20,30, 39, 43, 50, 60 and 88 KDa. RESULTS: The allergenic composition of spore and mycelium looked very similar. However, partial or total inhibition of the serum with a conidial or mycelial extract demonstrated that the total concentration of allergens (particulary 20 and 60 KDa molecules) was higher in the conidium than in the mycelium. CONCLUSIONS: These results indicated that conidium and mycelium contained the same allergenic determinants but at different concentration in the two propagule. Results with 50 % inhibited sera demonstrated also that the total concentration of allergens was higher in the conidium than in the mycelium.

Immunochemical comparison of the allergenic potency of spores and mycelium of cladosporium cladosporioides extracts by a nitrocellulose electroblotting technique / Comparación inmunoquímica de la potencia alergénica de los extractos de esporas y mecilio del cladosporium cladosporioides por una técnica de eletroblotting en nitrocelulosa

Background: The lack of well standardized or characterized extracts that contain the relevant allergens of the appropriate fungus is resulting in a high heterogeneity of the commercial preparation. Material and methods: Immunochemical detection of the allergens composition of spore and mycelium of C. cladosporioides was studied by electroblotting using sera from Cladosporium allergic patients and 125 I- anti- human IgE. A MW range of allergens between 16 to 88 KDa was identified. The most important with a MW of 16, 20,30, 39, 43, 50, 60 and 88 KDa. Results: The allergenic composition of spore and mycelium looked very similar. However, partial or total inhibition of the serum with a conidial or mycelial extract demonstrated that the total concentration of allergens (particulary 20 and 60 KDa molecules) was higher in the conidium than in the mycelium. Conclusions: These results indicated that conidium and mycelium contained the same allergenic determinants but at different concentration in the two propagule. Results with 50 % inhibited sera demonstrated also that the total concentration of allergens was higher in the conidium than in the mycelium

Antecedentes: La falta de extractos estandarizados o caracterizados que contienen alergenos relevantes de componentes fúngicos, da como resultado una elevada heterogeneidad de los preparados comerciales. Material and methods. La detección inmunoquímica de la composición alergénica de las esporas y del micelio de C. Cladosporioides ha sido estudiada por el método del electroblotting usando sueros procedentes de pacientes alérgicos a Cladosporium y anti-IgE humano marcado con Yodo 125. El peso molecular de los alergenos identificados fue situado entre 16 y 88 Kda. Los más importantes con un peso molecular de 16, 20, 30, 39, 43, 50, 60 y 88 Kda. Resultados: La composición alergénica detectada en las espora y el micelio ha sido idéntica. Sin embargo, la inhibición parcial o total del suero con extractos de esporas o micelios ha demostrado que la dosis de alergenos (especialmente de 20 y 60 KDa) ha sido más alta en las esporas que en el micelio. Conclusiones: Estos resultados indican que las esporas y los micelios contienen los mismos determinantes alergénicos pero a diferente concentración en ambos extractos. Con un 50% de inhibición sérica, la concentración de alergenos fue más elevada para las esporas que para los milecelios

Comparison of the allergenic potency of spores and mycelium of Cladosporium.

The allergenic potency of spore and mycelium extracts of Cladosporium was estimated by RAST, RAST inhibition and PCA tests. Spores contained a concentration of allergens higher than mycelia. Results of PCA tests suggested that spores contained specific allergens. However, in a comparative study of extracts from different species of Cladosporium animal and human models gave different estimates of the allergenic potency of the different species. In spite of these variations it was shown that extracts from spores of Cladosporium contained the highest amount of Cladosporium allergens.

Comparison of the allergenic potency of spores and mycelium of Cladosporium

The allergenic potency of spore and mycelium extracts of Cladosporium was estimated by RAST, RAST inhibition and PCA tests. Spores contained a concentration of allergens higher than mycelia. Results of PCA tests suggested that spores contained specific allergens. However, in a comparative study of extracts from different species of Cladosporium animal and human models gave different estimates of the allergenic potency of the different species. In spite of these variations it was shown that extracts from spores of Cladosporium contained the highest amount of Cladosporium allergens

El potencial alergénico de los extractos de las esporas y del micelio de Cladosporium ha sido valorado por los métodos de RAST, RAST inhibición y PCA. Las esporas contienen una dosis de alérgenos más elevada que el micelio. Los resultados del ensayo PCA sugieren que las esporas contienen alérgenos específicos. Sin embargo, en un estudio comparativo de los extractos procedentes de diferentes especies de Cladosporium, el modelo animal y humano han dado diferentes estimaciones del poder alérgenico entre las distintas especies. Aunque haya variaciones, se ha demostrado que los extractos de las esporas de Cladosporium contienen una cantidad más elevada de los alérgenos

[Neonatal screening of glucose-6-phosphate dehydrogenase deficiency in umbilical cord blood]. / Dépistage néonatal du déficit en glucose-6-phosphate déshydrogénase (G6PD) sur sang de cordon.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most frequent enzyme deficiency. It is a sex-linked genetic disease concerning mostly african, mediterranean and far-eastern populations. The main clinical expression is a hemolytic anemia which can be acute or chronic. During the neonatal period the disease may manifest as neonatal jaundice. We have been asked by the neonate department to set up a blood screening test for this deficiency. We have therefore developed a test using umbilical cord blood. The assay of G6PD has been automatised and red blood cell aspartate-amino-transferase (ASAT) chosen as a reference enzyme to evaluate the age of red blood cells. Normal values of G6PD, ASAT and G6PD/ASAT ratio have been calculated from 235 cord samples. Genetic frequency of this deficiency in 2002 was 6% in male and 1% in female newborns.

[An outbreak of allergy to horses in children. A review of 56 recent cases]. / Recrudescence de l'allergie au cheval chez l'enfant? A propos de 56 observations récentes.

Over the past 11 years, signs of allergy were observed in 56 children and adolescents in contact with horses. The cases consisted of 35 boys and 21 girls, 35 of them were under 10 years of age. The main clinical signs were ocular symptoms (36), asthma (30) and rhinopharyngitis (24). All the children had very positive cutaneous prick tests and specific IgE (class 3 and 4: 62%) and were polysensitized. In several children, the first manifestation occurred at the time of the first known contact with a horse or pony. No further contact was usually the only therapeutic solution. In disabled children, allergy to horses must be considered when clinical signs of allergy occur during therapeutic riding sessions.

[Does the allergic child become sensitized to small domestic mammals (guinea pig, hamster, rabbit?)]. / L'enfant allergique se sensibilise-t-il aux petits mammifères domestiques (cobaye, hamster, lapin)?

Our own works show among allergic children a frequency of sensitization of 30% to cats and of 17% to dogs. But, the proportion of sensitizations to other small domestic mammals such as guinea pigs, hamsters of rabbits seems to be unknown. Among atopic children with a known contact with those pets we noted: for guinea pig: 29% positive cutaneous or RAST tests 21% clinical signs in presence of animals; for hamster respectively 28% and 6.5%; for rabbit respectively 18% and 12.3%. Guinea pigs seem to be more sensitizing animals, followed by rabbits, then by hamsters. As for cats and dogs, eviction is imperative when a sensitization is demonstrated.

[Is it necessary to test children having summer respiratory problems with cereal pollens?]. / Faut-il tester aux pollens de céréales les enfants présentant des troubles repiratoires estivaux?

We tested 100 children with clinical signs that occurred in spring or summer. Prick tests were made with extracts of wild grass pollens (Dactylis, Phleum, Lolium) and with separate cereal pollens (Wheat, oats, Sweet-corn Barley and Rye). In 20 children whose test was negative for wild grass pollen, we saw no sensitization to cereals. In 80 children with grass pollenosis, Barley and Rye are the cereals that gave the most positive prick tests, whilst sweet corn is the cereal with most negative tests.

[Is immediate allergy the source of hematuria in children]. / L'allergie immédiate peut-elle être à l'origine d'hématuries chez l'enfant?

The authors report 2 cases of children presenting with episodes of hematuria, frequently following attacks of asthma or of allergic rhinitis. In both children, a respiratory provocation test with a mite extract triggered off hematuria. One child had an IgA nephropathy. In the second, the renal biopsy was normal and the cause of the hematuria remains unknown. These case reports suggest the need to carry out a systematic search for microscopic hematuria in patients with asthma.

[Chronic rhinopathies of childhood (apropos of 265 cases)]. / Les rhinopathies chroniques de l'enfant (à propos de 265 observations).

We observed among 265 children with a chronic rhinopathy: 187 allergic rhinopathies, associating sneezing, rhinorrhea and nasal eosinophilia, particularly caused by a sensitization for mites. 10 non allergic eosinophilic rhinopathies with identical clinical signs, and major nasal eosinophilia without any allergic factor. 68 non allergic non eosinophilic rhinopathies, where are preponderant nasal obstruction, olfaction troubles, ORL infections (otitis and sinusitis). Their etiology is probably various: psychic factors, ciliary dyskinesias, vasomotor phenomena...

[Comparative study of 3 positive controls for routine prick tests in children: 48/80, 1 mg histamine and 5 mg histamine]. / Etude comparative chez l'enfant de trois témoins positifs pour la pratique des prick tests: 48/80, histamine 1 mg et histamine 5 mg.

We compared, among 153 children 2 to 14 years old, responses to some positive control solutions for prick tests made with a Stallerpoint. We obtained the following values: -6% - 48/80:4.46 +/- 1.88 mm -1 mg/ml Histamine:3.49 +/- 1.67 mm -5 mg/ml Histamine:5.13 +/- 1.78 mm. 1 mg/ml histamine gives too many null responses (12/153) to represent a valuable positive control. 5 mg/ml histamine, on the other hand, gives some very strong reactions which may induce an underestimation of responses to allergen tests. Compound 48/80 seems to remain the best positive control for prick test exploration of immediate allergy in children. An alternative might be an intermediate solution of histamine 2.5 mg/ml.

[Long-term tolerance of specific hyposensitization with the aid of calcium phosphate-adsorbed mite extracts]. / Tolérance à long terme de l'hyposensibilisation spécifique à l'aide d'extraits d'acariens adsorbés sur phosphate de calcium. A propos de 90 observations traitées de façon semi-rapide.

90 children receive a specific hyposensitization with calcium phosphate adjuvant purified Dermatophagoides pteronyssinus allergens. We note 44% very good results, 37% good results and 19% failures. We propose to begin with 1/1,000,000 concentration every other day until 1/10,000 concentration is obtained under antihistamine protection. Tolerance is satisfactory: moderate respiratory reaction are pointed out for 22 patients. No anaphylactic shock is encountered. For 80 children with more than three years recall, we totalled 3,700 injections for 340 watching years. Only eight patients are sight lost. 1/10,000 concentration is obtained in 72/80 followed up children.

[Severe local reactions to mosquito bites. Apropos of 20 cases]. / Les réactions locales majeures aux piqûres de moustique. A propos de 21 observations.

We relate 21 observations of children who present major local reactions, immediate and delayed after mosquito bites. An associated reagin mechanism is obvious in 8 patients: five positive cutaneous or RAST tests and three a basophil degranulation tests. They are mostly young children (14/21 an less 5 years old). After some difficult years, the evolution is finally favourable with a specific hyposensitization or under antihistaminic protection (ketotifen) during summer.

[What problems does childhood toxocariasis currently pose? Apropos of 6 clinical cases]. / Quels problèmes posent actuellement les toxocaroses de l'enfant? (a propos de six observations cliniques).

About six observations of toxocariasis (visceral larva migrans syndrome). We relate six observations of toxocariasis among children. In one case, an ocular localization is probable. For other five patients, they are inapparent forms. The allergologist pediatrician may be consulted because of a major hypereosinophilia (greater than 10,000/mm3) and an elevation of total IgE (greater than 2,000 UI/ml). Allergic and current parasitologic assays are negative and diagnostic key is given by toxocara serology. We insist on interest and reliability of passive hemagglutination test with a purified antigen (titer greater than or equal to 1/320). Treatment now is preferably flubendazole (50 mg/kg/day for six days) eventually renewed.