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Recovery of complete genome sequences of Crimean-Congo haemorrhagic fever virus (CCHFV) directly from clinical samples: A comparative study between targeted enrichment and metagenomic approaches.
D'Addiego, Jake; Wand, Nadina; Afrough, Babak; Fletcher, Tom; Kurosaki, Yohei; Leblebicioglu, Hakan; Hewson, Roger.
Afiliação
  • D'Addiego J; UK Health Security Agency, Science Group, Porton Down, Salisbury, United Kingdom; Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom. Electronic address: jake.daddiego@ukhsa.gov.uk.
  • Wand N; UK Health Security Agency, Science Group, Porton Down, Salisbury, United Kingdom.
  • Afrough B; UK Health Security Agency, Science Group, Porton Down, Salisbury, United Kingdom.
  • Fletcher T; Department of Clinical Sciences, Liverpool School of Tropical Medicine, Liverpool, United Kingdom.
  • Kurosaki Y; National Research Centre for the Control and Prevention of Infectious Diseases, Nagasaki University, Japan.
  • Leblebicioglu H; VM Medicalpark Samsun Hospital, Samsun, Türkiye.
  • Hewson R; UK Health Security Agency, Science Group, Porton Down, Salisbury, United Kingdom; Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom; Department of Clinical Sciences, Liverpool School of Tropical Medicine, Liverpool, United Kingdom; Na
J Virol Methods ; 323: 114833, 2024 Jan.
Article em En | MEDLINE | ID: mdl-37879367
ABSTRACT
Crimean-Congo haemorrhagic fever (CCHF) is the most prevalent human tick-borne viral disease, endemic to the Balkans, Africa, Middle East and Asia. There are currently no licensed vaccines or effective antivirals against CCHF. CCHF virus (CCHFV) has a negative sense segmented tripartite RNA genome consisting of the small (S), medium (M) and large (L) segments. Depending on the segment utilised for genetic affiliation, there are up to 7 circulating lineages of CCHFV. The current lack of geographical representation of CCHFV sequences in various repositories highlights a requirement for increased CCHFV sequencing capabilities in endemic regions. We have optimised and established a multiplex PCR tiling methodology for the targeted enrichment of complete genomes of Europe 1 CCHFV lineage directly from clinical samples and compared its performance to a non-targeted enrichment approach on both short-read and long-read sequencing platforms. We have found a statistically significant increase in mapped viral sequencing reads produced with our targeted enrichment approach. This has allowed us to recover near complete S segment sequences and above 90% of the M and L segment sequences for samples with Ct values as high as 31.3. This study demonstrates the superiority of a targeted enrichment approach for recovery of CCHFV genomic sequences from samples with low virus titre. CCHFV is an important vector-borne human pathogen with wide geographical distribution. The validated methodology reported here adds value to front-line public health laboratories employing genomic sequencing for CCHFV Europe 1 lineage surveillance, particularly in the Balkan and Middle Eastern territories currently monitoring the spread of the pathogen. Tracking the genomic evolution of the virus across regions improves risk assessment and directly informs the development of diagnostics, therapeutics, and vaccines.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Vacinas / Vírus da Febre Hemorrágica da Crimeia-Congo / Febre Hemorrágica da Crimeia Limite: Humans Idioma: En Revista: J Virol Methods Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Vacinas / Vírus da Febre Hemorrágica da Crimeia-Congo / Febre Hemorrágica da Crimeia Limite: Humans Idioma: En Revista: J Virol Methods Ano de publicação: 2024 Tipo de documento: Article