A high-throughput DNA microarray platform for the diagnosis of viruses
transmitted by arthropods and rodents
Mohd Jaseem Khan1; Helda Liz Alfonso1; Amanda Cristina Trabuco1*; Soraya Jabur Badra2; Luiz Tadeu
Moraes Figueiredo2; Victor Hugo Aquino Quintana1
1Laboratório
de Virologia, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo.
2Centro de Pesquisa em Virologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo.
*[email protected]
Brazil is a tropical country where several viruses transmitted by
arthropods (arbovirus) and rodents (robovirus) are circulating.
Considering the public health significance of these viruses, there is a
great interest for the development of rapid diagnostic methods.
The DNA microarray platform has emerged recently as a highthroughput method for pathogen detection.
Based on 466 viruses sequences (roboviruses and arboviruses)
deposited in the GenBank, 4715 probes with 60 nucleotides were
designed (approximately 10 probes per virus).
The microarray slide (RoboArbovirusChip) was prepared by Agilent
(USA) in an eight array per slide format. Each array contained the
4715 probes with at least three replicates, plus positive and
negative controls. The DetectiV software1 was used to analyze the
fluorescence data.
Development of a DNA microarray platform for the detection of roboviruses
and arboviruses.
To evaluate the RoboArbovirusChip, 15 viruses were tested and detected. RoboArbovirusChip was also able to detect Dengue virus from serum
samples of infected patients. No cross-hybridization was observed with RNA obtained from C6/36 cells.
Fluorescence Intensity (Log2)
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Figure 2. Detection of 15 laboratory viruses from 4 families on RoboArbovirusChip: Flaviviridae –Dengue virus
type-1 (DENV-1), Dengue virus type-2 (DENV-2), Dengue virus type-3 (DENV-3), Dengue virus type-4 (DENV-4),
Saint Louis encephalitis virus (SLEV), Rocio virus (ROCV), Yellow fever virus (YFV), Bussuquara virus (BSQV),
Iguape virus (IGUV) and Ilheus virus (ILHV) – Alphaviridae – Mayaro virus (MAYV) – Rabdoviridae – Piry virus
(PIRV) – and Bunyaviridae – Oropouche virus (OROV), Rio Mamoré virus (RIOMV) and Guaroa virus (GROV).
C6/36: Experiment Negative Control. *: p<0,05 compared to slide Negative Control.
Fluorescence Intensity (Log2)
Figure 1. RoboArbovirusChip slide image obtained
from GenePix 4000B Microarray Scanner after
sample incubation.
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DENV-1
DENV-2
DENV-3
DENV-4
Negative
Control
Positive
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Figure 3. Detection of DENV-1, DENV-2, DENV-3 and DENV-4 from human serum samples.
*: p<0,05 compared to slide Negative Control.
RoboArbovirusChip was able to detect both laboratory virus and clinical virus isolates. These results strongly suggest the potential of this platform for
the used as a rapid diagnostic method for high-throughput monitoring of roboviruses and arboviruses
1. http://detectiv.sourceforge.net/
2. Palacios, G., et al. 2007. Emerg Infect Dis 13, 73-81.