Development of digital PCR molecular tests for clinical practice: principles, practical implementation and recommendations.

Jérôme Alexandre Denis; Juliette Nectoux; Pierre-Jean Lamy; Christelle Rouillac Le Sciellour; Hélène Guermouche; Anne-Sophie Alary; Olivier Kosmider; Nasrin Sarafan-Vasseur; Cécile Jovelet; Benoit Busser; Philippe Nizard; Valérie Taly; Frédéric Fina

Development of digital PCR molecular tests for clinical practice: principles, practical implementation and recommendations.

Jérôme Alexandre Denis ¹ Juliette Nectoux ² Pierre-Jean Lamy ³ Christelle Rouillac Le Sciellour ⁴ Hélène Guermouche ⁵ Anne-Sophie Alary ⁶ Olivier Kosmider ⁷ Nasrin Sarafan-Vasseur ⁸ Cécile Jovelet ⁹ Benoit Busser ¹⁰ Philippe Nizard ¹¹ Valérie Taly ¹¹ Frédéric Fina ¹²

1 I-STEM - Institut des cellules souches pour le traitement et l'étude des maladies monogéniques

2 Service de biochimie et de génétique moléculaire [CHU Cochin]

3 IRCM - U896 Inserm - UM1 - Institut de recherche en cancérologie de Montpellier

4 Fédération de génétique [CHI de Poissy-Saint-Germain en Laye]

5 Service d'immunologie et hématologies biologiques [Saint-Antoine]

6 CHRU Montpellier - Centre Hospitalier Régional Universitaire [Montpellier]

7 IC UM3 (UMR 8104 / U1016) - Institut Cochin

8 GPMCND - Génomique et Médecine Personnalisée du Cancer et des Maladies Neuropsychiatriques

9 Labo RT - Laboratoire de recherche translationnelle

AMMICa - Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse

10 UF Cancérologie biologique et biothérapie

11 MEPPOT - U1147 - Médecine Personnalisée, Pharmacogénomique, Optimisation Thérapeutique

12 AMU MED - Aix-Marseille Université - Faculté de médecine

Abstract : Digital PCR (dPCR) is a 3rd generation technology that complements traditional end-point PCR and real-time PCR. It was developed to overcome certain limitations of conventional amplification techniques, in particular for the detection of small amounts of nucleic acids and/or rare variants. This technology is in a full swing because of its high sensitivity and major applications in various domains such as oncology, transplantation or non-invasive prenatal testing. Consequently, PCRd also has great interest in many areas of medical biology, particularly for clinical applications aiming at detecting and quantifying specific genetic or epigenetic alterations of nucleic acids, even with specimens containing very low concentration of the nucleic acids of interest (e.g. liquid biopsies). However, this technique requires a good training of users and compliance with certain precautions. A lack in such a knowledge can lead to many errors in the conduct of the experiment and the interpretation of the results. In this review, we present the context in which this technology has emerged by describing in particular its principle and the main factors that can influence the quality of the analysis. Then, we propose a number of practical recommendations for the implementation of a test based on dPCR in clinical laboratories with an eye on quality requirements.

Keywords : digital PCR dropplet micro-compartiments microfluidics quantitative PCR real-time PCR recommendations

Type de document :

Article dans une revue

Domaine :

Sciences du Vivant [q-bio]

Liste complète des métadonnées

https://hal-normandie-univ.archives-ouvertes.fr/hal-02353393
Contributeur : Nasrin Vasseur <>
Soumis le : jeudi 7 novembre 2019 - 11:59:02
Dernière modification le : jeudi 14 novembre 2019 - 17:22:04

Development of digital PCR molecular tests for clinical practice: principles, practical implementation and recommendations.

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