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Problems in containing aflatoxins,Various alfatoxin quantification systems,ELISA: Ideal test for Sub-Saharan Africa,ELISA and data analysis,Quantification of aflatoxins in different commodities
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www.iita.org
P Lava Kumar and R Bandyopadhyay
Fostering Food Safety Monitoring in SSA
Synergistic interaction
with Hepatitis-B & C
Malnutrition
Liver cancer
Trade
Restrictions
Quality reductionAflatoxin
contamination
in food and feed
Reduced ability to
cope with diseases,
especially HIV/AIDS
Liver cirrhosis, immuno-suppression,
blocks nutrient absorption, growth
abnormalities, etc.
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• The most effective measures to control aflatoxins depends on
a program that “monitors” their presence in food and feeds.
But,
• Monitoring programs in SSA relies on importation of diagnostic
tools, which are expensive, some have short shelf-life and
prohibitive for majority of the stakeholders.
• “Sustainability” depends on the
-Sustainable access to basic reagents at affordable costs.
-Suitability for application in a minimally equipped labs.
Problems in containing aflatoxins
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Various alfatoxin
quantification systems
TLC HPLC LC-MS
Analytical Methods
Immuno-columns ELISA Lateral Flow Strips
Serological Methods
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• ELISA have been proven to be easy for adoption in developing
countries.
ELISA: Ideal test for SSA
• IITA has developed cheap and high-through put screening systems
which dramatically reduces the phenotyping cost.
Afla-ELISATM is a simple and low-cost
test for quantitative estimation of
aflatoxins developed at IITA.
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Components
-AFB1-BSA standards -Coating buffer concentrate
-AFB1 standards -Dilution buffer concentrate
-AFB1 polyclonal antibodies -Substrate buffer concentrate
-Sample preparation & ELISA protocol -Wash buffer concentrate
Afla-ELISA kit
Coast:
US$ 2 per sample
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Three simple steps
1. Sample Extraction
3. Data input into software
2. ELISA
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1. Ground 20 g sample (maize)
2. Resuspend in 100 ml of
0.5% KCL in 70% Methanol
3. Mix the suspension (30 min) 4. Filter the extract
Sample extraction
5. Use it in ELISA
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Measure in an ELISA plate
reader set with 405 nm
Final Reaction
Input data into XL program sheet
to estimate AFB1 concentration in
unknown samples
y = -2.1638x + 2.2869
R2 = 0.9953
-2.000
-1.000
0.000
1.000
2.000
3.000
0.000 0.500 1.000 1.500 2.000
ELISA and data analysis
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Blender
Orbital Shaker ELISA Plate reader
Pipette pH Meter ELISA Plates
General labware and a computer
Requirements for aflatoxin lab
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Quantification of aflatoxins in
different commodities
0.000
0.200
0.400
0.600
0.800
1.000
1.200
1.400
1.600
1.800
1 2 3 4 5 6 7 8 9
Log AfB1(ng/ml)
A4
05
nm
millet
sorghum
ground nut
cassava
rice
maize
Buffer
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Specificity of Afla-ELISA
0.0
20.0
40.0
60.0
80.0
100.0
120.0
1 2 3 4 5
LogAfB1(ng/ml)
% b
ind
ing
B1
B2
G1
G2
FB1
OTA
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Afla-ELISA
Trade Mark Registration
TM
Next step:
Product registration
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Next Steps and Exit Strategy
2. Product Development [OUTPUT]
1. Research
3. Product Deployment [OUTPUT]
Fostering utilization by NARS, researchers, monitoring agencies etc)
-Product registration - Accreditation - policy
-Training and facilitating infrastructure development
3. Utilization of Afla-ELISA for food safety monitoring [OUTCOME]
ExitBy 3rd party
By
IITA
Definite Exit
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Thank you
A&H Program R4D Week 2009