Variation in copy number of the 28S rDNA of Aspergillus fumigatus measured by droplet digital PCR and analog quantitative real-time PCR

Alexandre Alanio (1, 2, 3, 4) , Aude Sturny-Leclère (4, 1) , Marion Benabou (5) , Nicolas Guigue (3) , Stéphane Bretagne (3, 4, 1, 2)

1 CNRMA - Centre National de Référence des Mycoses invasives et antifongiques - Mycologie moléculaire
2 UPD7 - Université Paris Diderot - Paris 7
3 Laboratoire de Parasitologie-Mycologie [CHU Saint Louis, Paris]
4 Mycologie moléculaire
5 GHFC (UMR_3571 / U-Pasteur_1) - Génétique humaine et fonctions cognitives - Human Genetics and Cognitive Functions

Droplet digital PCR (ddPCR) after DNA digestion yielded a 28S rDNA copy number of 61 to 86 copies/genome when testing 10 unrelated Aspergillus fumigatus isolates, higher than with quantitative PCR. Unfortunately, ddPCR after DNA digestion did not improve the sensitivity of our PCR assay when testing serum patients with invasive aspergillosis.

Keywords: Aspergillus fumigatus; copy number variation; Droplet digital PCR; rDNA; Real-time quantitative PCR

Domains: Life Sciences [q-bio]; Life Sciences [q-bio]; Biochemistry, Molecular Biology; Molecular biology; Life Sciences [q-bio]; Microbiology and Parasitology; Mycology

Alexandre Alanio, Aude Sturny-Leclère, Marion Benabou, Nicolas Guigue, Stéphane Bretagne. Variation in copy number of the 28S rDNA of Aspergillus fumigatus measured by droplet digital PCR and analog quantitative real-time PCR. Journal of Microbiological Methods, 2016, 127, pp.160--163. ⟨10.1016/j.mimet.2016.06.015⟩. ⟨pasteur-01405204⟩