Detection of Aspergillus antigens in experimental invasive aspergillosis using a Biotin-Avidin Linked Sandwich ELISA (BALISA)

Shamim Abdul Samad, Jacinta Santhanam, Hamidah Yusoff

Research output: Contribution to journalArticle

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 × 106 and 1 × 107 conidia. However, antigenemia was detected in only 0%, 50% and 75% of NZW rabbits inoculated with 1 × 106, 1 × 107 and 1 × 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.

Original languageEnglish
Pages (from-to)3-13
Number of pages11
JournalAsia-Pacific Journal of Molecular Biology and Biotechnology
Volume12
Issue number1-2
Publication statusPublished - Dec 2004

Fingerprint

Aspergillosis
Avidin
Aspergillus
Antigens
Biotin
Enzyme-Linked Immunosorbent Assay
Rabbits
Water
Antibodies
Aspergillus fumigatus
Fungal Spores
Proteins
Serum
Concanavalin A
Organ Culture Techniques
Infection
Sensitivity and Specificity

Keywords

  • Antigen detection
  • Experimental invasive aspergillosis
  • Sandwich ELISA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Bioengineering

Cite this

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title = "Detection of Aspergillus antigens in experimental invasive aspergillosis using a Biotin-Avidin Linked Sandwich ELISA (BALISA)",
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 × 106 and 1 × 107 conidia. However, antigenemia was detected in only 0{\%}, 50{\%} and 75{\%} of NZW rabbits inoculated with 1 × 106, 1 × 107 and 1 × 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.",
keywords = "Antigen detection, Experimental invasive aspergillosis, Sandwich ELISA",
author = "Samad, {Shamim Abdul} and Jacinta Santhanam and Hamidah Yusoff",
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AU - Samad, Shamim Abdul

AU - Santhanam, Jacinta

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N2 - 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 × 106 and 1 × 107 conidia. However, antigenemia was detected in only 0%, 50% and 75% of NZW rabbits inoculated with 1 × 106, 1 × 107 and 1 × 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.

AB - 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 × 106 and 1 × 107 conidia. However, antigenemia was detected in only 0%, 50% and 75% of NZW rabbits inoculated with 1 × 106, 1 × 107 and 1 × 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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