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Development of a Membrane-Based Immunofiltration Assay for the Detection of T-2 Toxin

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Title Development of a Membrane-Based
Immunofiltration Assay for the Detection of T-2
Toxin
 
Creator Pal, Arindam
Acharya, Debopam
Saha, Debjani
Dhar, Tarun K
 
Subject Infectious Diseases and Immunology
 
Description An improved analytical device capable of performing
simultaneous immunofiltration-based immunoassay on
30 samples in the presence of reference standards has
been developed. The device consists of a rectangular
membrane with 36 antibody spotted zones, one end of
which was attached to a semirigid polyethylene card. A
piece of wetted filter paper between the membrane and
the polyethylene card absorbs the added reagent. The
assay is a competitive one using T-2 toxin-horseradish
peroxidase (T-2 toxin-HRP) as the labeled analyte and
4-chloro-1 naphthol (4CN) as the substrate. Signal amplification
was done by the Super-CARD signal amplification
method. Semiquantitative results were obtained by
visual comparison of the color intensity of a sample spot
with those of reference standards. Densitometric analysis
was used for quantitation. The method allows rapid and
easy determination of T-2 toxin in wheat and poultry feed
with detection limits of 12.5 and 25 Ìg kg-1, respectively,
with accuracy and precision. Matrix interference was
eliminated by appropriate dilution of sample extracts with
assay buffer. The detection sensitivity in ELISA was 10-
fold higher than that in the membrane-based method.
Noninfected samples were spiked with T-2 toxin at several
concentrations and analyzed by the present method and
rapid ELISA. Mean recoveries by both methods were
between 80 and 108%. The correlation between the two
methods was excellent (R2 ) 0.99).
 
Date 2004
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/466/1/ANALYTICAL_CHEMISTRY%2C_76(_14)%2C__4237%2D4240_[45].pdf
Pal, Arindam and Acharya, Debopam and Saha, Debjani and Dhar, Tarun K (2004) Development of a Membrane-Based Immunofiltration Assay for the Detection of T-2 Toxin. Analytical Chemistry., 76 (14). pp. 4237-4240.
 
Relation http://dx.doi.org/10.1021/ac049631s
http://www.eprints.iicb.res.in/466/