Prototype of oligonucleotide microarray for detection of pathogens relating to arena- and filoviridae families

Abstract

A prototype of oligonucleotide microarray for detection of Lassa, Junin, Machupo, Guanarito viruses (Arenaviridae family), Ebola and Marburg viruses (Filoviridae family) was presented. An original approach founded on virus proteins (nucleocapsid protein for Junin, Guanarito, Machupo viruses and RNA-dependent RNA-polymerase for Lassa, Ebola and Marburg viruses) amino acid sequences analysis with subsequent transform of revealed unique peptides into due sets of oligonucleotides was used to design probes for hybridization and primers.

An updated review and current challenges of Guanarito virus infection, Venezuelan hemorrhagic fever

“…GTOV, a BSL-4 pathogen, is included in group I of the priority classification by the World Health Organization because of its severity and often fatal outcome due to the lack of effective treatment and preventive measures. Thus, rapid laboratory diagnosis is very important and urgent for administering adequate treatment [ 38 ].…”

“…Although IgM can be detected rapidly, cross-reactions can occur between GTOV and other arenaviruses. Neutralizing antibodies, which are much more selective and would be ideal for early diagnosis, usually appear too late after infection [ 38 ]. Since GTOV and other arenaviruses are classified as BSL-4 pathogens, their diagnosis in laboratories without BSL-4 facilities is difficult, and thus, in order to overcome these difficulties, antibody detection by enzyme-linked-immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFA) using recombinant viral NPs may be an option, not as much for etiological diagnosis, but for seroepidemiological surveillance, because these tests can be performed in BSL-2 laboratories, which are more accessible [ 40 , 41 ].…”

“…However, due to the short duration of viremia, the usefulness of PCR can be limited. Furthermore, detection of neutralizing antibodies is difficult, as it also requires a BSL-4 laboratory, which is usually unavailable in endemic areas [ 38 ]. Multiplex real-time qRT-PCR and TaqMan qRT-PCR assays for the rapid detection of viruses causing hemorrhagic fever, including GTOV, have proved to be specific and sensitive [ 42 – 44 ].…”

“…In the last decade, the use of analytical microarrays has become a promising diagnostic method that significantly reduces the time needed for analysis by allowing parallel diagnosis, testing one sample with multiple probes that discriminate among different infectious agents. For example, in one study, a prototypic microarray was developed to distinguish members of the families Filoviridae and Arenaviridae , including GTOV, and this represents an important tool for the diagnosis of VHF [ 38 ]. Nonetheless, its cost and technical requirements limit its applicability in GTOV-endemic areas.…”