Michael Z. Zheng, John L.

Richard and Johann Binder

Presented by
Widiastuti Setyaningsih

Under Supervision by
Professor Armando
Durant
 INTRODUCTION
Mycotoxin Producer

Contamination

FOOD
BORNE
DESEASE
Mycotoxin Analysis
Conventional
Analysis Rapid Method
Analysis
Easier to use
Faster
Less expensive
Rapid Methods for Mycotoxin
Analysis
 The term ‘rapid method’ usually refers to a method much
faster than respective reference methods and also has a
tendency of promoting the method.

1. ELISA
2. Membrane based immunoassay
 Flow-through assay
 Lateral flow test
1. Flurometric Assay
 Immunoaffinity column clean-up
 SPE column clean-up
1. Fluorescent polarization
Antibody
 Enzyme linked immuno-sorbent
assay (ELISA)
(a) Sample mixed
with conjugate;
(b)Mixed content
added to antibody
coated well;
(c) Mycotoxin binds
to antibody in the
1st incubation;
(d)Unbound
materials are
rinsed away in the
washing step;
(e)Substrate is
added to develop
color;
(f) Stop solution is
added to stop the
• Direct Competitive ELISA
• Anti-mycotoxin anti-body is coated on a membrane
surface
• Mycotoxin and mycotoxin- enzyme conjugate
compete for the
limited antibody binding sites.
Membrane
based
immunoassay
: Flow-through
assay
Principle of membrane-based
flow through test
• (in the absence of toxin (left),
color is developed;
• in the presence of toxin (right),
color development is suppressed).
A typical immunochromatography test strip
• Sample pad,
• Conjugate pad,
• A membrane,
• An absorbent pad and
• An adhesive backing.

The competitive reaction scheme is used most

often when testing for small molecules with
single antigenic determinants such as
mycotoxins
Membrane
based
immunoassay :
Lateral flow test

Schematic illustration
of a lateral flow test.
(a) Configuration;
(b) Reagents placement.
contains anti-mycotoxin
antibody that is
immobilized onto
a solid support
such as agarose gel
in phosphate buffer,

all of which is contained

in a small plastic
cartridge
Principle of immunoaffinity
columns
Solid-phase extraction (SPE)
column clean-up
SPE columns or cartridges are
Silica gel,
C18 bonded to silica gel,
Forisil or ion exchange resins.

 A one-step SPE cleanup

Solid-phase extraction (SPE)
column clean-up

Principle of a one-step solid phase extraction

column
1. Evanescent wave technology
2. MIPs
3. Microarray technology
4. Luminex xMAPÒ technology
Evanescent wave technology
Surface plasmon resonance
biosensor
Principle of Surface Plasmon
Resonance (SPR)
L: light source,
D: photodiode array detector,
P: prism,
S: sensor surface,
F: flow cell.

(The two thick lines in the reflected

beam
projected on to the D symbolize the
dropping of light intensity following
the resonance phenomenon at time =
t1 and t2. The line projected at t1
corresponds to the situation before
binding of antigens to the antibodies
on the surface and t2 is the position
 Fiber Optic
Immunosensor

Principle of Fibre Optic Immunosensor

 Critical View
 Insufficient validation of methods causes the methods to be
limited to those matrices for which they were validated

 (ELISA) methods also need to be validated and developed that

will distinguish among the mycotoxins individually or
collectively (not only valid for each the matrix) since the
cross reactivity might occur.

 Even the method is quantitative or semi-quantitative,

confirmation analysis still need to be conducted.

 The microarray technology can be used not only for microbial

fungus determination as mentioned by the author, but also for
the simultaneous detection of mycotoxin (aflatoxin B1 and
 Critical View
 No conclusion that may mention the recommended rapid
method which is considered has better performance for
mycotoxin analysis and apart from MIPs, the author did not
describe how the methods have the possibility as potential
technologies to be able to apply for mycotoxin analysis

 The fluorometric assay with SPE clean-up cannot be

comparable since the analyte only aflatoxin B1 instead of total
aflatoxin (B1, B2 and G1).

 This review may helpful for the food producers or food

inspector institutions to choose the suitable method to
monitor the quality of the product apart from mycotoxin
contamination