The innovative molecular biology techniques
Constantly updating and using the latest molecular biology technologies to meet current and future challenges.
What do DNA-based studies involve?
In 1953, James Watson and Francis Crick discovered the double helix structure of deoxyribonucleic acid (DNA). This discovery revolutionized not only the scientific community but also our everyday lives.
DNA is the memory of a living being, serving as a code programming the development of cells and tissues, and provides a method of transmitting genes (reproduction, heredity).
The study of DNA allows:
- the identification of a person or a species
- the detection of anomalies or mutations
The study of DNA and other genetic information uses molecular biology tools permitting the collection of information about a living being at the molecular level (DNA).
Our growing knowledge on genetics and molecular biology over the last 20 years has permitted huge advances, including:
- new therapies in the fight against certain diseases
- detection of viruses such as SARS-CoV-2 during the Covid-19 pandemic.
New generation sequencing (NGS) is a DNA sequencing approach allowing the decoding of genetic information. This new innovative method can handle large amounts of information allowing ultra-high throughput at much higher speeds compared to traditional methods.
The standard technique based
on quantitative PCR (qPCR)
Also referred to as real-time PCR (RT-PCR), this technique remains the gold standard of all molecular biology methods in the field of diagnostics. It is reliable, sensitive, robust and can be used in high throughput.
The detection and quantification of microorganisms
Detection of bacteria, viruses or fungi in different matrices:
- plant
- soil
- wastewater
- cosmetic cream
- air samples
Case study
Since 2014, a qPCR method is used in the early detection of the agent responsible for powdery mildew of grapevines (Erysiphe necator). This method enables an early alert on the presence of powdery mildew before the appearance of symptoms. Since 2022 this method has been made easily accessible, in a ready-to-use kit in the idetectTM range, to laboratories able to perform molecular analyses.
Monitoring resistance to fungicides
Detection and quantification of genetic variants: for example, mutations of genes coding for proteins targeted by and thus offering resistance to fungicides
Case study
Resistance to quinone outside inhibitors (QoI), fungicides acting to block respiration, in agents responsible for downy mildew of grapevines is caused by a mutation in the cytochrome b gene (G143A). The frequency of this mutation is monitored in mixed populations to evaluate the level of resistance by qPCR.
The method of sequencing small fragments, pyrosequencing
Small fragments (<200pb) at a time are sequentially sequenced via enzymatic reactions. Following PCR-based amplification of the region of interest, the amplicons are sequenced by incorporation of labeled nucleotides.
The signal translated as nucleotide sequences is displayed in real time allowing the detection and quantification of genetic variations. With the PyroMark® 48 from Qiagen, pyrosequencing has received a new make-over and gained in speed, reliability and accessibility.
Case study
What are the limitations of qPCR? Difficulties can be encountered:
- When several mutations are present at the same site
- With unknown mutations
- With deletions or insertions of several nucleotides.
In these cases, pyrosequencing offers a more precise solution:
- SARS-CoV-2 in wastewater: detection and quantification of variants. To meet the challenge relating to the emergence of numerous variants of the virus, pyrosequencing ensures their efficient tracking.
- Resistance to fungicides: our team has discovered a new mechanism of resistance shown by the agent responsible for grapevine powdery mildew (Plasmopara viticola) to a fungicide called cyazofamid (cellular respiration inhibitor). This resistance comes from a 6 nucleotide insertion-induced modification of the fungicide target. The frequency of genes carrying this insertion can be quantified by pyrosequencing but not by qPCR.
Our solutions associated with the technology explained
idetect
Phytosanitary product testing
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