Development of a non-infectious control for viral hemorrhagic fever PCR assays.
PLoS Negl Trop Dis
; 18(4): e0011390, 2024 Apr.
Article
de En
| MEDLINE
| ID: mdl-38648254
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.
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Fièvres hémorragiques virales
Limites:
Humans
Langue:
En
Journal:
PLoS Negl Trop Dis
/
PloS neglected tropical diseases
/
Plos negl. trop. dis
Sujet du journal:
MEDICINA TROPICAL
Année:
2024
Type de document:
Article
Pays d'affiliation:
Nouvelle-Zélande
Pays de publication:
États-Unis d'Amérique