Development of a non-infectious control for viral hemorrhagic fever PCR assays.

Knox MA; Bromhead C; Hayman DTS

Development of a non-infectious control for viral hemorrhagic fever PCR assays.

dc.citation.issue	4
dc.citation.volume	18
dc.contributor.author	Knox MA
dc.contributor.author	Bromhead C
dc.contributor.author	Hayman DTS
dc.contributor.editor	Viennet E
dc.coverage.spatial	United States
dc.date.accessioned	2024-05-08T02:37:55Z
dc.date.available	2024-05-08T02:37:55Z
dc.date.issued	2024-04-22
dc.description.abstract	Assay validation is an essential component of disease surveillance testing, but can be problematic in settings where access to positive control material is limited and a safety risk for handlers. Here we describe a single non-infectious synthetic control that can help develop and validate the PCR based detection of the viral causes of Crimean-Congo hemorrhagic fever, Ebola virus disease, Lassa fever, Marburg virus disease and Rift Valley fever. We designed non-infectious synthetic DNA oligonucleotide sequences incorporating primer binding sites suitable for five assays, and a T7 promotor site which was used to transcribe the sequence. Transcribed RNA was used as template in a dilution series, extracted and amplified with RT-PCR and RT-qPCR to demonstrate successful recovery and determine limits of detection in a range of laboratory settings. Our results show this approach is adaptable to any diagnostic assay requiring validation of nucleic acid extraction and/or amplification, particularly where sourcing reliable, safe material for positive controls is infeasible.
dc.description.confidential	false
dc.format.pagination	e0011390-
dc.identifier.author-url	https://www.ncbi.nlm.nih.gov/pubmed/38648254
dc.identifier.citation	Knox MA, Bromhead C, Hayman DT. (2024). Development of a non-infectious control for viral hemorrhagic fever PCR assays.. PLoS Negl Trop Dis. 18. 4. (pp. e0011390-).
dc.identifier.doi	10.1371/journal.pntd.0011390
dc.identifier.eissn	1935-2735
dc.identifier.elements-type	journal-article
dc.identifier.issn	1935-2727
dc.identifier.number	0011390
dc.identifier.pii	PNTD-D-23-00631
dc.identifier.uri	https://mro.massey.ac.nz/handle/10179/69516
dc.language	eng
dc.publisher	PLOS
dc.publisher.uri	https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0011390
dc.relation.isPartOf	PLoS Negl Trop Dis
dc.rights	(c) 2024 The Author/s
dc.rights	CC BY 4.0
dc.rights.uri	https://creativecommons.org/licenses/by/4.0/
dc.title	Development of a non-infectious control for viral hemorrhagic fever PCR assays.
dc.type	Journal article
pubs.elements-id	488514
pubs.organisational-group	College of Health