The recombinant antigen ASPND1r from Aspergillus nidulans is specifically recognized by sera from patients with aspergilloma.
A 996 bp Aspergillus nidulans cDNA encoding the ASPND1 immunodominant antigen was cloned and expressed in Escherichia coli as a fusion protein with the enzyme glutathione S-transferase (GST) from Schistosoma japonicum. The GST-ASPND1 fusion protein was purified from isolated bacterial inclusion bodies by preparative SDS-PAGE. After cleavage with thrombin, the ASPND1 recombinant antigen (ASPND1r) and the GST protein were separated by SDS-PAGE and immunoblotted with a number of different human sera. The sera from 22 (88%) of 25 patients with an aspergilloma recognized the ASPND1r recombinant antigen on immunoblots. Forty-nine normal human sera and 14 sera from patients with other infections were unreactive. The ASPND1r expressed in E. coli could therefore be used, in combination with previously reported recombinant antigens, as a standardized antigen for serological and clinical diagnosis of Aspergillus-associated diseases.
Puente
Pilar
P
Departamento de Microbiología y Genética, Ed. Departamental Biología, Lab. 218, Paseo del Campo Charro s/n, Universidad de Salamanca, 37007 Salamanca, Spain.
Leal
Fernando
F
Departamento de Microbiología y Genética, Ed. Departamental Biología, Lab. 218, Paseo del Campo Charro s/n, Universidad de Salamanca, 37007 Salamanca, Spain.
D000595:Amino Acid Sequence; D000818:Animals; D000908:Antibodies, Fungal / Q000032:analysis / Q000276:immunology*; D000946:Antigens, Fungal; D001228:Aspergillosis / Q000097:blood / Q000235:genetics / Q000276:immunology*; D001233:Aspergillus nidulans / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D012331:RNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000302:isolation & purification; D012549:Schistosoma japonicum / Q000235:genetics; D017422:Sequence Analysis, DNA; D013917:Thrombin / Q000494:pharmacology
C105207:ASPND1 protein, Aspergillus nidulans; D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D004269:DNA, Bacterial; D018076:DNA, Complementary; D005656:Fungal Proteins; D006023:Glycoproteins; D012331:RNA, Fungal; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D013917:Thrombin