An innovative DNA analysis to control raw materials and finished products : Metabarcoding analysis

Fabienne MOREAUQualtech Group/ADNid Company

Qualtech group/ADNid

• ADNid is a provider of DNA analyses for the quality control of raw materials and finished products

– Research of Citrus reticulata in orange juice

– Research of other varieties in Alphonso Mango puree

Qualtech group/ADNid

• Qualtech group is a provider of analyses in agrofood field

– Physicochemical analyses

– Microbiology analyses

– Contaminants (heavy metal, pesticides..)

– Sensory analyses

Qualtech group/ADNid

• A full service for quality control of rawmaterial and finished products

• A unique know how in DNA extraction

• Innovative analyses « METABARCODING »

Next Generation Sequencing

• Technological leap to analyse the genome

• New and many sequences in databases

• New know how for ADNid: Bioinformaticanalysis

• New tool to control raw materials and finishedproducts

DNA extraction

Degree of the DNA alteration- +

Abondant and

intact DNA

Traces

of damaged DNA

Degree of the technological process

Finished productRaw material Intermediate material

Damaged

DNA

MetabarcodingDNA EXTRACTION

RARE AND DAMAGED DNA

PCR AMPLIFICATIONUNIVERSAL OR

SPECIFIC TARGETS

- Species identification- (vegetable, fruit ….meat, fish, insect, microalgae)

- Microorganism screening - (bacteria, archaebacteria, yeast, fungi)

Specificities of the Metabarcoding

• Analysis of raw material or finished productwithout a priori

• Analysis in mixture

• Analysis of all the species of a material in one experiment

Species/Genus identification in multifruit juice and purée

• Experimental juiceExperimental composition DNA results

Apple Malus domestica

Orange Citrus

Banana Musa

Apricot Prunus armeniaca

The results could be the name of the species or the name of the genus

Species/Genus identification in multifruit juice

Room temperature Cold temperature

Day 0 One sample per bottle for fruit species identification and microorganism screeningDay 3 and 6 One sample per bottle for microorganism screening

Storage at 4°C afteropening

1 2

Species identification in multifruit juiceExperimental protocol

DNA EXTRACTIONRARE AND DAMAGED DNA

PCR AMPLIFICATIONWITH SPECIFIC

TARGETS OF VEGETABLE

Results for species identification

Expected composition

Found

Ananas Ananas comosus

apple Malus domestica

banana Musa

Passion fruit Passiflora

grape Vitis vinifera

JUICE 1Expected composition

Found

Ananas Ananas comosus

apple Malus domestica

banana Musa

Passion fruit Passiflora

Apricot Prunus Armeniaca

Orange Citrus

Papaya Carica papaya

Mango Mangifera indica

JUICE 2

Results for species identification

• Composition of the 2 juices was validated by the Metabarcoding analysis

• Taxonomic rank : specie or genus

• Sequences found in the sample are blasted with database in order to determine the genus or specie name

• Qualitative, not quantitative

Microorganism screeningExperimental protocol

DNA EXTRACTIONRARE AND DAMAGED DNA

PCR AMPLIFICATIONWITH SPECIFIC

TARGETS OF BACTERIA

Results for the Microbiom

Day 0 Day 3 Day 6

Around 10 different populations of bacteriaYeastFungi

Metabarcoding Microbiom

• Alive or dead microorganism

• Unculturable microorganism

• Picture of the different populations

• Qualitative, Not quantitative

Applications of Metabarcoding- Target identification(meat, fish, insect, vegetables)- Microorganism screening (bacteria, archaebacteria, yeast,

fungi)- Validation of the specification- Process monitoring - Non compliance sample analyses - Supplier control : raw material, intermediary product, end-

product - Communication to consumer

Thank you

Fabienne Moreau

Scientific Director

Fabienne.moreau@qualtech-groupe.com

+ 33 6 14 25 07 44

You’re welcome