A DNA Microarray-Based Assay to Detect Dual Infection with Two Dengue Virus Serotypes

Abstract

Here; we have described and tested a microarray based-method for the screening of dengue virus (DENV) serotypes. This DNA microarray assay is specific and sensitive and can detect dual infections with two dengue virus serotypes and single-serotype infections. Other methodologies may underestimate samples containing more than one serotype. This technology can be used to discriminate between the four DENV serotypes. Single-stranded DNA targets were covalently attached to glass slides and hybridised with specific labelled probes. DENV isolates and dengue samples were used to evaluate microarray performance. Our results demonstrate that the probes hybridized specifically to DENV serotypes; with no detection of unspecific signals. This finding provides evidence that specific probes can effectively identify single and double infections in DENV samples.

Molecular and Immunological Diagnostic Techniques of Medical Viruses

“…DNA microarray was also used to identify viral causes of meningitis and encephalitis with overall sensitivity of 93% and specificity of 100%, compared to single-virus PCR [ 104 ]. DNA microarray was also utilized for high-throughput multiplex detection of gastrointestinal viruses [ 105 ], viruses transmitted by small mammals and arthropods [ 106 ], herpesviruses, enteroviruses and flaviviruses [ 107 ], HIV-1, HIV-2, and hepatitis viruses [ 108 ] and dual infection with two dengue virus serotypes [ 109 ] in human specimens. DNA microarray was used to identify and genotype drug-resistant mutations of HIV [ 110 , 111 ] to detect and genotype drug-resistant hepatitis B virus (HBV) mutations [ 112 ], to detect and genotype SARS coronavirus [ 113 ], and to detect and determine lineage of influenza B viruses [ 114 ].…”

An intermolecular-split G-quadruplex DNAzyme sensor for dengue virus detection

“…By applying the equation of DL ¼ 3s b /slope (where s b ¼ the standard deviation of the blank), the detection limits of the serotype-specic assays were determined to range from 4.9 nM to 9.3 nM, i.e. DENV-1: Table 3 Absorbance readings generated by the multi-probe cocktail assay in the presence of various concentrations of different DENV serotype target strands Table 4 Comparative advantages and disadvantages of the split G4 DNAzyme-based assay with DNA microarrays, real time reverse-transcription PCR (qRT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP) and semi-quantitative nested PCR DENV serotyping assay Split G4 DNAzymebased assay 50 DNA microarray 51 qRT-PCR 52,53 RT-LAMP 54 8.8 nM, DENV-2: 4.9 nM, DENV-3: 9.3 nM and DENV-4: 5.1 nM. The difference in the detection limits obtained may be inuenced by the different hybridization strengths of the threestrand complex.…”

Challenges and Opportunities of Laboratory diagnosis of Dengue virus infection: a review

“…Application of microarray platforms in the diagnosis of dengue infection has been reported in several studies. Bumgarner and others demonstrated the simultaneous detection of dengue virus serotypes (DENV-1, DENV-2, DENV-3, and DENV-4) using complementary DNA based microarray [108]. Another study by Cretch et al (2014) showed the detection of anti-dengue immunoglobulin M antibodies using single-molecule array [109] and envelope domain III (EDIII) enzyme-linked immunosorbent assay-based array with an improved limit of detection compared to conventional IgM antibody capture enzyme-linked immunosorbent assay [110].…”