Fiche publication
Date publication
juin 2025
Journal
MethodsX
Auteurs
Membres identifiés du Cancéropôle Est :
Pr ROHR Olivier
Tous les auteurs :
Chaqroun A, El Soufi G, Gerber Z, Loutreul J, Cluzel N, Challant J, Delafoy D, Sandron F, Di Jorio L, Raffestin S, Maréchal V, Gantzer C, Olaso R, Deleuze JF, Rohr O, Boudaud N, Wallet C, Bertrand I
Lien Pubmed
Résumé
This study aimed to evaluate whether a nucleic acid extraction protocol specifically designed for raw wastewater (WW) provides a measurable advantage over protocols not originally intended for WW matrices. Three laboratories have independently compared two RNA extraction protocols, using paired WW samples. A common WW-designed protocol (Z), based on silica columns was tested across all labs. As comparators, three non-WW designed protocols were used: NS1 and NS2 both silica beads-based, with NS2 including an additional phenol-chloroform step; and SB a homemade protocol using also silica beads but differing in its formulation. As different samples were used across labs, direct statistical comparison was difficult; instead, paired comparisons within each lab were used to rank SARS-CoV-2 RNA detection and RT-qPCR inhibitor removal. NS1 yielded significantly higher SARS-CoV-2 RNA concentrations than Z (Logrank test, ≤ 0.001), though with RT-qPCR inhibition in one sample. NS2 also showed higher SARS-CoV-2 RNA detection than Z (Wilcoxon test, < 0.0001), with both protocols showing complete inhibitor removal. SB performed worse than Z for SARS-CoV-2 RNA detection (Logrank test, ≤ 0.05), and showed inhibition in one sample. NS2 was the most effective option for both RNA detection and inhibitor removal.
Mots clés
NS2, PCR inhibitors, RNA extraction, SARS-CoV-2, Silica beads, Silica columns, Wastewater
Référence
MethodsX. 2025 06;14:103323
