Immunoassays for detection of Mucor, Rhizopus and related genuses and speciei: development and applications

Mucor pusillus (Ag824) and Rhizopus nigricans extracts were used as the source of immunogen to immunize rabbits.

Rabbit antiserum against Mucor (AS824) was further enriched by affinity chromatography on immobilized antigen. Affinity purified antibodies (AS824 A) are available for research.

ELISA kit (K824) was constructed from AS824 antiserum as single site inhibitory assay, see IFU

Cross-reactivity of various fungal extracts has shown low to absent reactivity in K824 Mucor ELISA kit (see table below), except for Rhizopus extract which has shown identical reactivity. Therefore, the Mucor ELISA kit can be described as the kit for detection of all Mucorales family

Fungal extracts, 10 ug/ml by total protein	OD	Inhibition/X reactivity
Buffer	1,145
Aspergillus niger	1,195	NO
Aspergillus flavus	1,200	NO
Rhizopus nigricans	0,515	+++strong
Mucor pusillus	0,477	+++POS control
Phoma betae	1,264	NO
Alternaria alternate	1,221	NO
Fusarium moniliforme	1,185	NO

Conidia, and NOT the mycelium of the fungi is the sole source of the antigen detected by K824 Mucor ELISA kit (data not shown)

Accumulation of conidial antigens measured by ELISA in cultivation medium correlates well with the number of conidia (see figure below)

Affinity antibody against Mucor was used for immunocytochemical staining of fungal culture The pictures (see below) confirm the localization of target antigen in conidia.
Control staining (no anti-Mucor antibodies)

Specific staining (anti-Mucor antibodies)