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. 2020 Mar 28;7(4):ofaa111.
doi: 10.1093/ofid/ofaa111. eCollection 2020 Apr.

Contact Transmission of Vaccinia to an Infant Diagnosed by Viral Culture and Metagenomic Sequencing

Rebekah M Martin  1   2 , Kristina Burke  3 , Dinesh Verma  4 , Heng Xie  5 , Janine Langer  2 , Robert Schlaberg  1   5 , Sankar Swaminathan  4 , Kimberly E Hanson  1   2   4
Affiliations

Contact Transmission of Vaccinia to an Infant Diagnosed by Viral Culture and Metagenomic Sequencing

Rebekah M Martin et al. Open Forum Infect Dis. .

Abstract

Targeted molecular diagnostic tests and accurate immunoassays have transformed the landscape of clinical virology, calling into question the usefulness of traditional viral culture. Here we present a case where viral culture, followed by metagenomic sequencing, was central to the diagnosis of an unexpected viral infection, with significant clinical and public health implications.

Keywords: metagenomic sequencing; poxvirus; vaccine-transmitted; vaccinia; viral culture.

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Figures

Figure 1.
Figure 1.
Skin lesions. Ulcerative lesions in the buttock area ranging in size from 1 to 2.5 cm. Consent to use these images for publication was obtained from the patient’s family.
Figure 2.
Figure 2.
Identification and characterization of vaccinia virus from tissue culture by shotgun metagenomic RNA and DNA sequencing. A, Shotgun RNA sequencing revealed 2.8% of sequencing reads to be of viral origin (pie chart), resulting in a mean 126-fold coverage of the vaccinia virus genome. B, Ninety-two percent of shotgun DNA sequencing reads were of viral origin, producing >4000-fold mean coverage and allowing for construction of a high-quality viral consensus genome for the patient strain covering 199 077 nucleotides (99.92% of the reference genome). Vaccinia virus strain Acambis clone 2000, used in the second-generation smallpox vaccine (ACAM2000, GenBank accession number AY313847), was the most similar strain in public databases. Only a single nucleotide polymorphism (49676G>A, numbering from AY313847) relative to ACAM2000 was identified.
Figure 3.
Figure 3.
Transmission electron micrograph of MRC-5 cells infected with vaccinia virus. Representative images of vaccinia virus are shown at different magnifications (11 000×, 30 000×, and 67 000×). The large (~200 × 300 nm) brick-shaped virions with biconcave (dumbbell-shaped) electron-dense cores containing the viral genome are characteristic of poxviruses.
See this image and copyright information in PMC

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