ABSTRACT
Objectives: Precise HDV-RNA detection and quantification are pivotal for diagnosis and monitoring of response to newly approved treatment. We evaluate the performance of three HDV RNA detection and quantification assays. Methods: Hepatitis Delta RT-PCR system kit, EurobioPlex HDV assay, and RoboGene HDV RNA Quantification kit 2.0 were used for testing 151 HBsAg-positive samples, 90 HDV-RNA negative and 61 HDV-RNA positive. We also evaluated serial dilutions of the WHO international standard for HDV, PEI 7657/12. All HDV-RNA positive samples were genotyped using a next-generation sequencing strategy. Results: Qualitative results indicated a 100% concordance between tests. Quantitative results correlated well, r2 = 0.703 (Vircell-vs-Eurobio), r2 = 0.833 (Vircell-vs-RoboGene), r2 = 0.835 (Robogene-vs-Eurobio). Bias index was 2.083 (Vircell-vs-Eurobio), -1.283 (Vircell-vs-RoboGene), and -3.36 (Robogene-vs-Eurobio). Using the WHO IS, Vircell overestimated the viral load by 0.98 log IU/mL, Eurobio by 1.46 log IU/mL, and RoboGene underestimated it by 0.98 log IU/mL. Fifty-nine samples were successfully genotyped (Genotype 1, n=52; Genotype 5, n=7; Genotype 6, n=1), with similar results for correlation and bias. Conclusion: This study underscores the necessity of using reliable HDV-RNA detection and quantification assays, as evidenced by the high concordance rates in qualitative detection and the observed variability in quantitative results. These findings highlight the importance of consistent assay use in clinical practice to ensure accurate diagnosis and effective treatment monitoring of HDV infection.
Subject(s)
Genotype , Hepatitis D , Hepatitis Delta Virus , RNA, Viral , Viral Load , Hepatitis Delta Virus/genetics , Hepatitis Delta Virus/isolation & purification , Humans , RNA, Viral/genetics , Viral Load/methods , Hepatitis D/diagnosis , Hepatitis D/virology , Reagent Kits, Diagnostic/standards , Sensitivity and Specificity , High-Throughput Nucleotide Sequencing/methods , Molecular Diagnostic Techniques/methodsABSTRACT
INTRODUCTION: To characterize a carbapenem-resistant Enterobacter cloacae complex isolate recovered from a patient from Ukraine. METHODS: The isolate was sent to a regional reference laboratory for molecular characterization by whole genome sequencing. Susceptibility assays, carbapenemase identification, imipenem hydrolysis and clonality were performed. RESULTS: The isolate showed resistance or reduced susceptibility to all β-lactam agents tested. Genome analysis led to the identification of an NDM-1-producing E. cloacae complex strain that was assigned to a new multilocus sequence type, ST932. The blaNDM-1 enzyme was located in a conjugative IncX3 plasmid of ca. 50kb. In addition, blaCMH-3, a recently described AmpC β-lactamase sequence, which has not previously been reported in Europe, was also detected and its genetic environment was studied. CONCLUSION: To our knowledge, this is the first reported case in Europe of an E. cloacae complex strain that produces both blaNDM-1 and blaCMH-3
INTRODUCCIÓN: Caracterizar una cepa de Enterobacter cloacae complex resistente a los carbapenémicos detectado en un paciente procedente de Ucrania. MÉTODOS: Se envió la cepa a un laboratorio regional de referencia para proceder a su caracterización molecular mediante secuenciación del genoma completo. Se realizaron estudios de susceptibilidad, identificación de carbapenemasas, hidrólisis del imipenem y de clonalidad. RESULTADOS: El aislado mostró resistencia o sensibilidad reducida a todos los betalactámicos estudiados. El análisis molecular permitió la identificación de una cepa de E. cloacae complex productor de NDM-1 que se asignó a un nuevo secuenciotipo, ST932. Se localizó la enzima blaNDM-1 en plásmido conjugativo IncX3 de 50kb. Además, se detectó la enzima blaCMH-3, una nueva betalactamasa de tipo AmpC descrita recientemente pero no detectada con anterioridad en Europa, llevándose a cabo el estudio de su entorno genético. CONCLUSIÓN: Según nuestros conocimientos, esta es la primera descripción una cepa de E. cloacae complex productora de blaNDM-1 junto con blaCMH-3 en Europa
Subject(s)
Humans , Enterobacter cloacae/genetics , Enterobacter cloacae/drug effects , Carbapenem-Resistant Enterobacteriaceae/genetics , Enterobacteriaceae Infections/diagnosis , Enterobacteriaceae Infections/microbiology , Whole Genome Sequencing , Enterobacter cloacae/enzymology , Carbapenem-Resistant Enterobacteriaceae/enzymology , beta-Lactamases , Anti-Infective Agents/pharmacology , Microbial Sensitivity Tests , Spain , UkraineABSTRACT
INTRODUCTION: To characterize a carbapenem-resistant Enterobacter cloacae complex isolate recovered from a patient from Ukraine. METHODS: The isolate was sent to a regional reference laboratory for molecular characterization by whole genome sequencing. Susceptibility assays, carbapenemase identification, imipenem hydrolysis and clonality were performed. RESULTS: The isolate showed resistance or reduced susceptibility to all ß-lactam agents tested. Genome analysis led to the identification of an NDM-1-producing E. cloacae complex strain that was assigned to a new multilocus sequence type, ST932. The blaNDM-1 enzyme was located in a conjugative IncX3 plasmid of ca. 50kb. In addition, blaCMH-3, a recently described AmpC ß-lactamase sequence, which has not previously been reported in Europe, was also detected and its genetic environment was studied. CONCLUSION: To our knowledge, this is the first reported case in Europe of an E. cloacae complex strain that produces both blaNDM-1 and blaCMH-3.