Development and analytical validation of a group-specific RT-qPCR assay for the detection of the Simbu serogroup orthobunyaviruses
| dc.contributor.author | Camarão, A.A.R. | |
| dc.contributor.author | Swanepoel, Robert | |
| dc.contributor.author | Boinas, F. | |
| dc.contributor.author | Quan, Melvyn | |
| dc.contributor.email | melvyn.quan@up.ac.za | en_ZA |
| dc.date.accessioned | 2019-06-18T12:54:57Z | |
| dc.date.issued | 2019-09 | |
| dc.description.abstract | The Simbu serogroup within the genus Orthobunyavirus belongs to the family Peribunyaviridae and comprises 32 recognised three-segmented negative-sense single-stranded RNA viruses, with a cosmopolitan distribution. This group of arthropod-borne viruses includes important pathogens of humans and domestic animals e.g. Oropouche orthobunyavirus and Schmallenberg virus. Sensitive and specific diagnostic tools are required for recognition and control of outbreaks. A novel TaqMan® RT-qPCR assay was developed, optimised and analytically validated for the broad detection of the Simbu serogroup orthobunyaviruses. A region in the S segment, which encodes the nucleocapsid protein, was used to design a group primer set and a pair of differently labelled TaqMan® minor groove binder probes to distinguish phylogenetic clade A and B of the serogroup. Efficiencies determined for seven members of the group were 99% for Akabane orthobunyavirus (AKAV), 96% for Simbu orthobunyavirus (SIMV), 96% for Shuni orthobunyavirus (SHUV), 97% for Sathuperi orthobunyavirus (SATV), 84% for Shamonda orthobunyavirus (SHAV), 93% for Ingwavuma virus (INGV, now classified as Manzanilla orthobunyavirus) and 110% for Sabo virus (SABOV, now classified as AKAV). The 95% limit of detection (TCID50/reaction) was 10-3.61 for AKAV, 10-2.38 for SIMV, 10-3.42 for SHUV, 10-3.32 | en_ZA |
| dc.description.department | Veterinary Tropical Diseases | en_ZA |
| dc.description.embargo | 2020-09-01 | |
| dc.description.librarian | hj2019 | en_ZA |
| dc.description.uri | http://www.elsevier.com/locate/jviromet | en_ZA |
| dc.identifier.citation | Camarao AAR, Swanepoel R, Boinas F, Quan M, Development and analytical validation of a group-specific RT-qPCR assay for the detection of the Simbu serogroup orthobunyaviruses, Journal of Virological Methods (2019) 271: 113685, https://doi.org/10.1016/j.jviromet.2019.113685. | en_ZA |
| dc.identifier.issn | 0166-0934 (print) | |
| dc.identifier.issn | 1879-0984 (online) | |
| dc.identifier.other | 10.1016/j.jviromet.2019.113685 | |
| dc.identifier.uri | http://hdl.handle.net/2263/70230 | |
| dc.language.iso | en | en_ZA |
| dc.publisher | Elsevier | en_ZA |
| dc.rights | © 2019 Elsevier B.V. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Journal of Virological Methods. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. A definitive version was subsequently published in Journal of Virological Methods, vol. 271, art. 113685, pp. 1-10, 2019. doi : 10.1016/j.jviromet.2019.113685. | en_ZA |
| dc.subject | Simbu serogroup viruses | en_ZA |
| dc.subject | Clade A | en_ZA |
| dc.subject | Clade B | en_ZA |
| dc.subject | TaqMan® | en_ZA |
| dc.subject | Real-time RT-PCR | en_ZA |
| dc.subject | Orthobunyaviruses | en_ZA |
| dc.title | Development and analytical validation of a group-specific RT-qPCR assay for the detection of the Simbu serogroup orthobunyaviruses | en_ZA |
| dc.type | Postprint Article | en_ZA |