Allergens of Epicoccum nigrum grown in different media for quality source material.
BACKGROUND
The Epicoccum nigrum (EN) extract used in allergy disorders exhibits batch-to-batch variations in protein composition and allergenic potency. In this study, the allergens of EN grown in different media were investigated.


METHODS
EN was grown in five different nutrient media as stationary cultures at 25 degrees C for 5-23 days. The growth pattern was characterized by measuring dry weight, protein and carbohydrate content. The antigenic and allergenic content of EN extract was evaluated with EN-positive patients' sera and antibodies raised in rabbit.


RESULTS
The growth of EN in Czapeck Dox medium yielded insufficient material, while Sabouraud's broth with yeast extract (SBY) gave maximum spore-mycelial mass and protein content. Potato dextrose broth (PDB) and potato dextrose agar (PDA) showed higher dry weight and protein in 7-9-day cultures. SDS-PAGE resolved 26, 22, and 21 protein bands in EN extracts from cultures of day-13 SBY, day-7 PDB, and day-9 PDA, respectively. IgE/IgG immunoblots showed more allergenic (25)/antigenic (25) bands in EN cultured in SBY than in the others. Specific IgE ELISA and intradermal tests showed EN extract from day-13 culture in SBY to be the most potent.


CONCLUSIONS
The day-13 culture of EN in SBY was the most potent and may be selected for preparing EN extracts for diagnosis of allergy and future studies.
Bisht
V
V
Centre for Biochemical Technology, Delhi University Campus, India.
Singh
B P
BP
Arora
N
N
Sridhara
S
S
Gaur
S N
SN
D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000032:analysis / Q000276:immunology*; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000032:analysis / Q000276:immunology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000032:analysis*; D007074:Immunoglobulin G / Q000032:analysis*; D007428:Intradermal Tests; D003904:Mitosporic Fungi / Q000737:chemistry / Q000254:growth & development / Q000276:immunology*; D011817:Rabbits; D012130:Respiratory Hypersensitivity / Q000276:immunology*
D000485:Allergens; D002241:Carbohydrates; D003470:Culture Media; D005656:Fungal Proteins; D007074:Immunoglobulin G; D007073:Immunoglobulin E