Resequencing microarray method for molecular diagnosis of human arboviral diseases
Autor(es): Berthet N, Paulous S, Coffey L L, Frenkiel M P, Moltini I, Tran C, Matheus S, Ottone C, Ungeheuer M N, Renaudat C, Caro V, Dussart P, Gessain A, Desprès P
Resumo: Resequencing DNA microarray (RMA) technology uses probes designed to identify a panel of viral sequences. It can be used for detecting emerging viruses by revealing the nucleotide polymorphisms within the target of interest. As a new tool for molecular diagnosis of arbovirus infection, high density PathogenID v2.0 RMA (PID2-RMA) was assessed for the detection - genetic analysis of dengue, West Nile, - Chikungunya viruses in spiked blood samples or sera from individuals infected with dengue virus. Viral RNAs extracted from biological samples were retrotranscribed into cDNA - amplified using the Phi 29 polymerase-based method. This amplified cDNA was used for hybridization on PID2-RMA. A good specificity of RMA-based detection was demonstrated using a panel of arboviruses including Dengue, West Nile - Chikungunya viruses. This technology was also efficient for the detection - genetic analysis of the different serotypes of dengue virus in sera of infected patients. Furthermore, the mixing of dengue, West Nile - Chikungunya prototype viruses within a single sample of human blood did not interfere with the sensitivity of PID2-RMA. Our data show that high density PID2-RMA was suitable for the identification of medically important arboviruses. It appears to be particularly adapted to the genetic analysis of dengue, West Nile, - Chikungunya viruses in urgent clinical situations where the rapid identification - characterization of the pathogen is essential.
Palavras-Chave: Resequencing microarray; Arboviruses; Diagnosis; Human blood samples
Imprenta: Journal of Clinical Virology, v. 56, n. 3, p. 238-243, 2013
Identificador do objeto digital: 10.1016/j.jcv.2012.10.022
Descritores: Chikungunya virus - Arbovirus ; Chikungunya virus - DNA ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya Virus - Virus ; Chikungunya virus - Molecular screening ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Dengue ; Chikungunya virus - Epidemiology
Data de publicação: 2013
