Complete nucleotide sequence of Middelburg virus, isolated from the spleen of a horse with severe clinical disease in Zimbabwe.
Abstract: 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.
Publication Date: 2007-10-20 PubMed ID: 17947533DOI: 10.1099/vir.0.83076-0Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The article presents the complete genetic sequence of Middelburg virus (MIDV) strain MIDV-857, discovered in Zimbabwe. The virus was isolated from a horse that died due to severe clinical illness, signaling that the virus could be potentially harmful to equine species.
Isolation and Sequencing of Middelburg Virus
- The MIDV-857 strain was isolated from a horse in Zimbabwe in 1993. The horse exhibited serious clinical signs, leading to its death, which acted as the first evidence of MIDV’s potential to cause severe disease in equids (horses, donkeys, and zebras).
- The researchers successfully created full-length cDNA copies of the MIDV genome using an innovative Reverse Transcription Polymerase Chain Reaction (RT-PCR) process. This involved an ‘anchor primer’ coupled with the SMART protocol previously used to synthesize full-length cDNA from genome segments of double-stranded RNA viruses.
Genome Characteristics of MIDV
- The total genetic sequence or genome of MIDV-857 is 11,674 nucleotides long excluding the 5′-terminal cap structure and polyadenylate (poly(A)) tail, which varies from approximately 180 to 220 residues.
- The genome organization of MIDV is similar to other members of the Alphavirus family. It includes a read-through stop codon found between non-structural protein 3 (nsP3) and nsP4 genes.
Potential Recombination and Novel Features in MIDV Genome
- Phylogenetic analyses of MIDV’s structural protein amino acid sequences suggested a potential recombination event. The E1 gene of MIDV, in particular, appears to have been generated through recombination with a virus similar to the Semliki Forest virus.
- This hypothesis gained further support from bootscanning analysis deploying a recombination-detection program.
- The 3′ untranslated region of the MIDV-857 genome possesses a 112 nucleotide duplication, which is another unique characteristic of the virus’ genome.
Significance of Study and Final Remarks
- The study marks a landmark in MIDV research by reporting the first full-length sequence of MIDV, demonstrating potential recombination events and unique genetic features of the virus.
- They obtained the full-length sequence of MIDV from a single RT-PCR product, showing the efficiency and reliability of their RT-PCR technique.
Cite This Article
APA
Attoui H, Sailleau C, Mohd Jaafar F, Belhouchet M, Biagini P, Cantaloube JF, de Micco P, Mertens P, Zientara S.
(2007).
Complete nucleotide sequence of Middelburg virus, isolated from the spleen of a horse with severe clinical disease in Zimbabwe.
J Gen Virol, 88(Pt 11), 3078-3088.
https://doi.org/10.1099/vir.0.83076-0 Publication
Researcher Affiliations
- Department of Arbovirology, Institute for Animal Health, Pirbright, Woking, Surrey GU24 0NF, UK.
- Agence Française de Sécurité Sanitaire des Aliments, 22 rue Pierre Curie, 94703 Maisons-Alfort Cedex 07, France.
- Department of Arbovirology, Institute for Animal Health, Pirbright, Woking, Surrey GU24 0NF, UK.
- Unité de Virologie Moléculaire, Etablissement Français du Sang Alpes-Méditerranée, 149 Boulevard Baille, 13005 Marseille, France.
- Unité de Virologie Moléculaire, Etablissement Français du Sang Alpes-Méditerranée, 149 Boulevard Baille, 13005 Marseille, France.
- Unité de Virologie Moléculaire, Etablissement Français du Sang Alpes-Méditerranée, 149 Boulevard Baille, 13005 Marseille, France.
- Unité de Virologie Moléculaire, Etablissement Français du Sang Alpes-Méditerranée, 149 Boulevard Baille, 13005 Marseille, France.
- Department of Arbovirology, Institute for Animal Health, Pirbright, Woking, Surrey GU24 0NF, UK.
- Agence Française de Sécurité Sanitaire des Aliments, 22 rue Pierre Curie, 94703 Maisons-Alfort Cedex 07, France.
MeSH Terms
- 3' Untranslated Regions / genetics
- Alphavirus / genetics
- Alphavirus / isolation & purification
- Alphavirus Infections / veterinary
- Animals
- Base Sequence
- Codon, Terminator / genetics
- Genome, Viral
- Horse Diseases / virology
- Horses
- Molecular Sequence Data
- Nucleic Acid Conformation
- Phylogeny
- RNA Caps / genetics
- RNA, Messenger / genetics
- Recombination, Genetic
- Semliki forest virus / genetics
- Sequence Analysis, DNA
- Sequence Homology, Amino Acid
- Spleen / virology
- Viral Nonstructural Proteins / genetics
- Viral Structural Proteins / genetics
- Zimbabwe
Citations
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