What does 1% Horse DNA Mean

Testing for Foreign DNA

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

WHAT is FOREIGN DNA

DNA is the genetic print of life.
It is transferred from one generation to other carrying vital
information of life.
Has been widely used in Food testing procedures to detect
pathogens, species identifications and determining GM
content.
DNA can spread to unrelated species by either
naturally (antibiotic resistance among microorganisms) or
human interventions (during food processing, creation of
hybrids, introduction of gene to enhance quality and
quantity of product)

The latter is the foreign DNA

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

FOREIGN DNA in FOOD

Humans are continually exposed to DNA in
Inhaled organisms,

From a broad variety of food sources

Via microorganisms normally present.

Infectious agents entering the body

Humans ingest 0.1 g to 1 g of DNA per day (Deorfler 2000).

0.1% to 1% of dietary DNA is absorbed from the

gastrointestinal tract (Nielsen et al; 2005a,2006)

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

FOREIGN DNA in FOOD

FOOD AUTHENTICATION and SPECIES IDENTIFICATION
Food authentication and food safety are a

growing concern.
Food fraud is a universal problem to tackle.
Mislabeling of Food
Consumers are cheated by paying for a
species that they are not actually getting.

Fraud hampers conservation efforts (especially

in sea food items).
Enhances illegal trade activities

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

IMPLICATIONS OF FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

Food adulteration or contamination can lead to

Physiological Effect
Can have health impacts when species are riddled
with contaminants.
Significant public health issues will arise
Toxic to sensitive consumers.
Allergens can get introduced into our health system.

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

IMPLICATIONS OF FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

Food adulteration or contamination can lead to

Economical Effects
Detrimental to business some companies get
benefited.
The horsemeat scandal in Europe served as a strong
reminder of how damaging unreliable suppliers can
be to corporate brands.
It robs hard-earned profits from honest suppliers and
businesses through unfair competition.

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

IMPLICATIONS OF FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION
LETS TRY

Escolar

Nile Perch

Mako Shark

Rock Fish

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

IMPLICATIONS OF FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

Food adulteration or contamination can lead to

Social and Ethical effects
the impacts of fraud can be social and ethical
concerns.
Widespread public fear
Loss of public confidence in safety of food
Effectiveness of authorities is questioned?

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

IMPLICATIONS OF FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

Food adulteration or contamination can lead to

Religious
Presence of certain contaminants is not accepted by
religious beliefs.
Pork has zero tolerance in the region.
Certain Food items are not eaten in other parts of the
world.

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

PRESENCE OF HORSE DNA - 1%

Indicates that the food is contaminated with horse meat
First reported by FSAI 15th January
Followed by FSA UK performing screening tests and identifying

products that had presence of horse and pork meat.

Subsequently other reports emerged and a legal framework was
designed.
1% is the minimum threshold level for DNA or protein resulting
from the DNA present in the food product above which labeling
is mandatory

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

PRESENCE OF HORSE DNA - 1%

The new UK meat testing regime agreed by the Food Standards Agency,
retailers and processors will allow for a 1% tolerance level for the DNA of other
species in meat products.

At the meeting, the industry also agreed to share its own meat DNA test results
with the FSA.

FSA is carrying out its own food authenticity programme on processed meat
lines.

It is understood the 1% level will apply to both the industrys own tests and the
FSA authenticity programme.

The FSA and industry are confident that labs can reliably test to 1% and that
levels of DNA at sub 1% are unlikely to indicate deliberate contamination,

We will be looking at this issue further and engaging directly with consumers as
we move forward.

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

DNA testing has the answer

Accurately detect the presence or
absence.
Quantify to meet the regulations of

allowable limits.
Identify different species present in

processed food.

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

Polymerase Chain Reaction (PCR)

REAL TIME PCR

DNA SEQUENCING

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA - PCR

FOOD AUTHENTICATION and SPECIES IDENTIFICATION
Denaturation

ds DNA denatured
by heating >94oC

Extension

Annealing

1 PCR
cycle

Taq polymerase
extends target
sequences

Oligo primers bind

to target sequences

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA Real Time PCR

FOOD AUTHENTICATION and SPECIES IDENTIFICATION
3
3
5
Forward Primer

R
5

Q
Reverse Primer
3
5
3

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA - PCR

FOOD AUTHENTICATION and SPECIES IDENTIFICATION
DNA based testing for Species Identification

2013

600 + total Samples

4000+ total Samples

2012

2008

2005

Food Authentication and Species

Identification

Water and Food-borne Pathogen

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA - PCR

FOOD AUTHENTICATION and SPECIES IDENTIFICATION
DNA based testing for Horse Species Identification

2013

200+ total Samples

1 Positive

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA DNA SEQUENCING

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

DNA sequencing Procedures

1.
2.
3.
4.
5.
6.

7.

DNA Extraction & Quantitation

DNA Amplification
PCR product purification
Cycle sequencing
Cycle sequencing product
clean up
Amplified DNA product
Separation using Capillary
Electrophoresis
Sequence Analysis

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA DNA SEQUENCING

FOOD AUTHENTICATION and SPECIES IDENTIFICATION
DNA Sequencing based Identifications

2013

300+ total Samples

4000+ total Samples

2012

2008

2005

Food Authentication and Species

Identification

Water and Food-borne Pathogen

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

DNA Barcoding CO1 Sequencing BOLD

Unique sequence that assists in Species identification
Developed by Canadian Centre for DNA Barcoding

NCBI Genebank Database

Amplified and sequenced Species specific DNA sequences

can be matched and identified from the database.

FDA sea food Authentication

Recently USFDA has developed protocol for Seafood

identification based on mitochondrial CO1 sequences.

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

Species Identification by BOLD system

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION

Species Identification by NCBI system

www.ncbi.nih.gov/blast
DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

TESTING FOREIGN DNA

FOOD AUTHENTICATION and SPECIES IDENTIFICATION
DNA testing can solve many a problems for Food
authentication.

Choose the right technique depending on the

complexity of the sample and also the results required.
Establish a Food screening program for all stakeholders

in Food industry and ensure integrity of the product and

consumer interest.
Dubai Municipality Proactive Approach
One of the few Municipalities in the world that supports and

propagates DNA based testing services.

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra

THANK YOU ALL

ISO 17025 Accredited Laboratory

LB-62-Test

306, Riqa Al Buteen Plaza

Al Maktoum Road
Dubai
Tel: 00971 4 2270004
Fax: 00971 4 2270005

DIFSC 2013 | Advance Biotechnology Centre, Dubai | Dr. Sanjeet Mishra