Detection of Aspergillus Antigens in Experimental Invasive Aspergillosis
using a Biotin-Avidin Linked Sandwich ELISA (BALISA)
Shamim Abdul Samad1*, Jacinta Santhanam2, Hamidah Yusoff1
1Department of Medical Microbiology & Immunology, Faculty of Medicine, Universiti Kebangsaan Malaysia,
Cheras, 56000 Kuala Lumpur, Malaysia
2Department of Biomedical Sciences, Faculty of Allied Health Sciences, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysia
*Author for Correspondence.
Dept. Medical
Microbiology & Immunology, Faculty of Medicine, UKM
Jalan Yaacob Latiff, 56000 Kuala Lumpur, Malaysia.
Tel: 603-91702208; Fax: 603-91737336
Email: shamim@mail.hukm.ukm.my
Key words: Antigen detection, experimental invasive aspergillosis, sandwich ELISA
Abstract.
A biotin-avidin amplified sandwich ELISA utilizing polyclonal antibodies to water soluble (WS) mycelial antigens
of Aspergillus fumigatus was used to detect antigens in sera of rabbits with experimental invasive aspergillosis. Tricolor rabbits
were more susceptible to infection than NZW rabbits as evidenced by a faster progression of infection and greater isolation
rates of A. fumigatus from organ culture. Antigenemia was detected in all 11 Tricolor rabbits inoculated with 1 x 106 and 1 x 107
conidia. However, antigenemia was detected in only 0%, 50% and 75% of NZW rabbits inoculated with 1 x 106, 1 x 107 and
1 x 108 conidia, respectively. Tricolor rabbits demonstrated antigenemia in 75% (18 of 24) of sera, whilst only 36% (5 of 14)
of sera of NZW rabbits tested positive. The overall sensitivity and specificity of antigen detection in Tricolor and NZW
rabbits were 84.2% (16 of 19 rabbits) and 85.7% (12 of 14) respectively. The protein rich Concanavalin A unbound fraction
of WS antigens demonstrated immunoreactivity with the BALISA. Immunoblot studies using the capture antibody demonstrated
3 strongly immunoreactive regions of 38-46kD, 66-68kD and 73-85kD for WS antigens. The results of partial characterization
of WS antigens, suggests that the antigens detected are protein in nature.
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