RT Journal Article SR Electronic(1) A1 Attoui, Houssam A1 Sailleau, Corinne A1 Mohd Jaafar, Fauziah A1 Belhouchet, Mourad A1 Biagini, Philippe A1 Cantaloube, Jean François A1 de Micco, Philippe A1 Mertens, Peter A1 Zientara, StephanYR 2007 T1 Complete nucleotide sequence of Middelburg virus, isolated from the spleen of a horse with severe clinical disease in Zimbabwe JF Journal of General Virology, VO 88 IS 11 SP 3078 OP 3088 DO https://doi.org/10.1099/vir.0.83076-0 PB Microbiology Society, SN 1465-2099, AB The complete nucleotide sequence of Middelburg virus (MIDV) was determined for strain MIDV-857 from Zimbabwe. The isolation of this virus in 1993 from a horse that died showing severe clinical signs represents the first indication that MIDV can cause severe disease in equids. Full-length cDNA copies of the viral genome were successfully synthesized by an innovative RT-PCR amplification approach using an ‘anchor primer’ combined with the SMART methodology described previously for the synthesis of full-length cDNA copies from genome segments of dsRNA viruses. The MIDV-857 genome is 11 674 nt, excluding the 5′-terminal cap structure and poly(A) tail (which varies in length from approximately 180 to approximately 220 residues). The organization of the genome is like that of other alphaviruses, including a read-through stop codon between the nsP3 and nsP4 genes. However, phylogenetic analyses of the structural protein amino acid sequences suggested that the MIDV E1 gene was generated by recombination with a Semliki Forest virus-like virus. This hypothesis was supported by bootscanning analysis using a recombination-detection program. The 3′ untranslated region of MIDV-857 also contains a 112 nt duplication. This study reports the first full-length sequence of MIDV, which was obtained from a single RT-PCR product., UL https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.83076-0