Comparison of Biosafety and Diagnostic Utility of Biosample Collection Cards

Viruses. 2022 Oct 29;14(11):2392. doi: 10.3390/v14112392.

Abstract

Six different biosample collection cards, often collectively referred to as FTA (Flinders Technology Associates) cards, were compared for their ability to inactivate viruses and stabilize viral nucleic acid for molecular testing. The cards were tested with bluetongue virus, foot-and-mouth disease virus (FMDV), small ruminant morbillivirus (peste des petits ruminants virus), and lumpy skin disease virus (LSDV), encompassing non-enveloped and enveloped representatives of viruses with double-stranded and single-stranded RNA genomes, as well as an enveloped DNA virus. The cards were loaded with virus-containing cell culture supernatant and tested after one day, one week, and one month. The inactivation of the RNA viruses was successful for the majority of the cards and filters. Most of them completely inactivated the viruses within one day or one week at the latest, but the inactivation of LSDV presented a greater challenge. Three of the six cards inactivated LSDV within one day, but the others did not achieve this even after an incubation period of 30 days. Differences between the cards were also evident in the stabilization of nucleic acid. The amount of detectable viral genome on the cards remained approximately constant for all viruses and cards over an incubation period of one month. With some cards, however, a bigger loss of detectable nucleic acid compared with a directly extracted sample was observed. Using FMDV, it was confirmed that the material applied to the cards was sufficiently conserved to allow detailed molecular characterization by sequencing. Furthermore, it was possible to successfully recover infectious FMDV by chemical transfection from some cards, confirming the preservation of full-length RNAs.

Keywords: Ahlstrom-Munksjö; Copan; FTA (Flinders Technology Associates) cards; Lipofectamine 3000; Macherey-Nagel; Whatman; biosample collection cards; real-time RT-qPCR; sequencing; transfection; virus isolation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Containment of Biohazards
  • Foot-and-Mouth Disease Virus* / genetics
  • Peste-des-petits-ruminants virus* / genetics
  • RNA, Viral / genetics

Substances

  • RNA, Viral

Grants and funding

This study was funded by Macherey-Nagel (Düren, Germany), grant number Ri-0638. This grant Ri-0638 is a bi-lateral industrial contract between the Friedrich-Loeffler-Institut and Macherey-Nagel. However, the funding had no direct relation to the presented study. The sponsor may not influence the study design, the data analyses and publication of the research results in any way.