This document discusses the development of affordable tools for monitoring aflatoxin contamination in food in sub-Saharan Africa. It notes that current monitoring relies on expensive imported tools. The document then summarizes the development of Afla-ELISA, a simple, low-cost ELISA test for quantifying aflatoxins. It describes the three simple steps to use Afla-ELISA involving sample extraction, performing the ELISA test, and analyzing the data. The document concludes by outlining next steps to register and deploy the product to facilitate its use in food safety monitoring.

1. Fostering Food Safety Monitoring in SSA
P Lava Kumar and R Bandyopadhyay
Reduced ability to
Malnutrition
cope with diseases,
Quality reduction especially HIV/AIDS
Aflatoxin Liver cirrhosis, immuno-suppression,
contamination blocks nutrient absorption, growth
Trade in food and feed abnormalities, etc.
Restrictions
Synergistic interaction
Liver cancer with Hepatitis-B & C
www.iita.org

2. Problems in containing aflatoxins
• 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.
www.iita.org

3. Various alfatoxin
quantification systems
Analytical Methods
TLC HPLC LC-MS
Serological Methods
Immuno-columns ELISA Lateral Flow Strips
www.iita.org

4. ELISA: Ideal test for SSA
• ELISA have been proven to be easy for adoption in developing
countries.
Afla-ELISATM is a simple and low-cost
test for quantitative estimation of
aflatoxins developed at IITA.
• IITA has developed cheap and high-through put screening systems
which dramatically reduces the phenotyping cost.
www.iita.org

5. Afla-ELISA kit
Components
-AFB1-BSA standards -Coating buffer concentrate Coast:
-AFB1 standards -Dilution buffer concentrate US$ 2 per sample
-AFB1 polyclonal antibodies -Substrate buffer concentrate
-Sample preparation & ELISA protocol -Wash buffer concentrate
www.iita.org

6. Three simple steps
1. Sample Extraction
2. ELISA
3. Data input into software
www.iita.org

7. Sample extraction
3. Mix the suspension (30 min) 4. Filter the extract
1. Ground 20 g sample (maize)
2. Resuspend in 100 ml of
0.5% KCL in 70% Methanol 5. Use it in ELISA
www.iita.org

8. ELISA and data analysis
Final Reaction Measure in an ELISA plate
reader set with 405 nm
3.000
y = -2.1638x + 2.2869
2.000 2
R = 0.9953
1.000
Input data into XL program sheet
0.000
0.000 0.500 1.000 1.500 2.000
to estimate AFB1 concentration in
-1.000 unknown samples
-2.000
www.iita.org

9. Requirements for aflatoxin lab
Orbital Shaker ELISA Plate reader
Blender
Pipette pH Meter ELISA Plates
General labware and a computer
www.iita.org

10. Quantification of aflatoxins in
different commodities
millet
1.800 sorghum
1.600 ground nut
1.400 cassava
1.200 rice
A405nm
1.000 maize
0.800 Buffer
0.600
0.400
0.200
0.000
1 2 3 4 5 6 7 8 9
Log AfB1(ng/ml)
www.iita.org

11. Specificity of Afla-ELISA
120.0
100.0
80.0 B1
% binding
B2
G1
60.0
G2
FB1
40.0 OTA
20.0
0.0
1 2 3 4 5
LogAfB1(ng/ml)
www.iita.org

12. Trade Mark Registration
TM
Afla-ELISA
Next step:
Product registration
www.iita.org

13. Next Steps and Exit Strategy
1. Research 
2. Product Development [OUTPUT] 
Exit
IITA
By
By 3rd party
3. Product Deployment [OUTPUT]
Fostering utilization by NARS, researchers, monitoring agencies etc)
-Product registration - Accreditation - policy
-Training and facilitating infrastructure development
Definite Exit
3. Utilization of Afla-ELISA for food safety monitoring [OUTCOME]
www.iita.org

14. Thank you
A&H Program R4D Week 2009 www.iita.org