Detection and discrimination of orthopoxviruses using microarrays of immobilized oligonucleotides

Real-Time PCR to Identify Variola Virus or Other Human Pathogenic Orthopox Viruses

“…Although these methods provide high specificity and allow species and sometimes strain differentiation, they are very labor-intensive and time-consuming and thus may delay diagnosis. Newer approaches that use speciesspecific oligonucleotide hybridization on a microchip (24 ) and fluorescence resonance energy transfer probes on the LightCycler instrument (25,26 ) have also been described. Another approach, the 5Ј nuclease PCR or the TaqMan assay, first developed by Holland et al (27 ) and improved by Lee et al (28 ), allows for the simultaneous amplification and detection of nucleic acids in real time.…”

Application of a Microarray-Based Assay for the Study of Genetic Diversity of West Nile Virus

“…It also may allow for the detection of those mutations even when they occur at relatively low levels (up to 1%) as in the case of mixtures of quasispecies that cannot be detected by traditional direct sequencing methods (Cherkasova et al, 2003;Leberre at al., 2007). The efficiency of microarrays for identification and discrimination of closely related bacteria and viruses has been previously demonstrated Hsia et al, 2007;Laassri et al, 2003;Nordström et al, 2005;Volokhov et al, 2002;Wade et al, 2004). The use of oligonucleotide microchips for screening of random mutations is based on the ability of microarrays to identify the presence of singlenucleotide mutations in the hybridization template (Hacia et al, 1999;Urakawa et al, 2003).…”

“…The microarrays consisting of multiple individual short oligoprobes were shown to be an efficient and sensitive genetic method for detection of single point mutations in viral and bacterial genomes Grinev et al, 2008b;Laassri et al, 2003Laassri et al, , 2005Laassri et al, , 2007Volokhov et al, 2002). Microarray assays can also help simultaneously detect and identify the genotype and strain of common food-borne viruses without using PCR (Chen et al, 2011).…”

Smallpox vaccines: New formulations and revised strategies for vaccination

“…Clear discrimination between MPXV and other orthopoxviruses is possible by the use of nucleic acid amplification technologies that detect specific differences in the genomic DNA sequence of the viruses. [16][17][18][19][20] The human infection dose of MPXV is not known. Yet, human monkeypox is believed to result from either respiratory, percutaneous or permucosal exposures.…”