Improvement in detecting cytomegalovirus drug resistance mutations in solid organ transplant recipients with suspected resistance using next generation sequencing

López-Aladid, RubénGuiu, AlbaMosquera Gutierrez, Maria del MarLópez-Medrano, FranciscoCofán, FredericLinares, LauraTorre-Cisneros, JulianVidal, ElisaMoreno, AsunciónAguado, José MaríaCordero, ElisaMartin-Gandul, CeciliaCarratalà, JordiSabé, NuriaNiubó, JordiCervera, CarlosCapón, AliciaCervilla, AnnaSantos, MartaBodro, MartaMuñoz, PatriciaFariñas, María CarmenAntón, AndrésAranzamendi, MaitaneMontejo, MiguelPerez-Romero, PilarLen, OscarMarcos, María Ángeles2019-11-252019-11-252019PLoS One. 2019 Jul 18;14(7):e0219701.1932-6203http://hdl.handle.net/20.500.12105/8705OBJETIVES: The aim of this study was to identify CMV drug resistance mutations (DRM) in solid organ transplant (SOT) recipients with suspected resistance comparing next-generation sequencing (NGS) with Sanger sequencing and assessing risk factors and the clinical impact of resistance. METHODS: Using Sanger sequencing as the reference method, we prospectively assessed the ability of NGS to detect CMV DRM in the UL97 and UL54 genes in a nationwide observational study from September 2013 to August 2016. RESULTS: Among 44 patients recruited, 14 DRM were detected by Sanger in 12 patients (27%) and 20 DRM were detected by NGS, in 16 (36%). NGS confirmed all the DRM detected by Sanger. The additional six mutations detected by NGS were present in <20% of the sequenced population, being located in the UL97 gene and conferring high-level resistance to ganciclovir. The presence of DRM by NGS was associated with lung transplantation (p = 0.050), the administration of prophylaxis (p = 0.039), a higher mean time between transplantation and suspicion of resistance (p = 0.038) and longer antiviral treatment duration before suspicion (p = 0.024). However, the latter was the only factor independently associated with the presence of DRM by NGS in the multivariate analysis (OR 2.24, 95% CI 1.03 to 4.87). CONCLUSIONS: NGS showed a higher yield than Sanger sequencing for detecting CMV resistance mutations in SOT recipients. The presence of DRM detected by NGS was independently associated with longer antiviral treatment.engVoRhttp://creativecommons.org/licenses/by/4.0/Improvement in detecting cytomegalovirus drug resistance mutations in solid organ transplant recipients with suspected resistance using next generation sequencingAtribución 4.0 Internacional31318908147e021970110.1371/journal.pone.02197011932-6203PloS oneopen access