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Long-Range Polymerase Chain Reaction Method for Sequencing the Ebola Virus Genome From Ecological and Clinical Samples.

ABSTRACT: Sequencing viral genomes during an outbreak can facilitate response and containment efforts. In this study, we describe a reverse transcription long-range polymerase chain reaction for efficient amplification and sequencing of the Ebola virus (EBOV) genome in 2 seminested reactions. We demonstrate that our method remains robust with complex biological samples by amplifying and sequencing the EBOV genome from EBOV-infected nonhuman primates (NHPs). We further demonstrate that we are able to recover viral genomes from starting concentrations as low as 103 50% tissue culture infective dose (TCID50)/mL, suggesting that this method can be employed to sequence EBOV genomes from ecologically or clinically derived samples.

SUBMITTER: Seifert SN

PROVIDER: S-EPMC6249577 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature

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Long-Range Polymerase Chain Reaction Method for Sequencing the Ebola Virus Genome From Ecological and Clinical Samples.

Seifert Stephanie N SN Schulz Jonathan E JE Matson M Jeremiah MJ Bushmaker Trenton T Marzi Andrea A Munster Vincent J VJ

The Journal of infectious diseases 20181101 suppl_5

Sequencing viral genomes during an outbreak can facilitate response and containment efforts. In this study, we describe a reverse transcription long-range polymerase chain reaction for efficient amplification and sequencing of the Ebola virus (EBOV) genome in 2 seminested reactions. We demonstrate that our method remains robust with complex biological samples by amplifying and sequencing the EBOV genome from EBOV-infected nonhuman primates (NHPs). We further demonstrate that we are able to recov ...[more]

PMID: 30085166

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