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.

www.iita.org

• 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

www.iita.org

Various alfatoxin

quantification systems

TLC HPLC LC-MS

Analytical Methods

Immuno-columns ELISA Lateral Flow Strips

Serological Methods

www.iita.org

• 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.

www.iita.org

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

www.iita.org

Three simple steps

1. Sample Extraction

3. Data input into software

2. ELISA

www.iita.org

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

www.iita.org

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

www.iita.org

Blender

Orbital Shaker ELISA Plate reader

Pipette pH Meter ELISA Plates

General labware and a computer

Requirements for aflatoxin lab

www.iita.org

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

www.iita.org

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

www.iita.org

Afla-ELISA

Trade Mark Registration

TM

Next step:

Product registration

www.iita.org

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

www.iita.org

Thank you

A&H Program R4D Week 2009