A new multiplex RT-qPCR method for the simultaneous detection and discrimination of Zika and chikungunya viruses
| Autor: | Marie-Alice Fraiture, Myrielle Dupont-Rouzeyrol, Vanessa Suin, Linda Garlant, Jessica Vanhomwegen, Sylvia Broeders, Els Vandermassen, Dominique Rousset, Steven Van Gucht, Nancy H. C. Roosens |
|---|---|
| Přispěvatelé: | Sciensano [Bruxelles], Réseau International des Instituts Pasteur (RIIP), Cellule d'Intervention Biologique d'Urgence - Laboratory for Urgent Response to Biological Threats (CIBU), Institut Pasteur [Paris], Institut Pasteur de Nouvelle-Calédonie, Institut Pasteur de la Guyane, This work was supported by the project ORIENT-EXPRESS financed by the WIV-ISP(presently named Sciensano, RP-PJ project number 0000754) and partially by the Arbo-VIRTUESS project financed by the Actions Concertées Interpasteuriennes(project number ACIP 2014-053)., Institut Pasteur [Paris] (IP) |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Identification viruses MESH: Chikungunya Fever medicine.disease_cause Genome law.invention Zika virus Disease Outbreaks 0302 clinical medicine law [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases Discrimination Multiplex 030212 general & internal medicine Chikungunya MESH: Disease Outbreaks Polymerase chain reaction [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases biology MESH: Multiplex Polymerase Chain Reaction MESH: Real-Time Polymerase Chain Reaction Zika Virus Infection virus diseases General Medicine 3. Good health Infectious Diseases [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology Chikungunya virus Microbiology (medical) 030106 microbiology MESH: Zika Virus Computational biology Real-Time Polymerase Chain Reaction Sensitivity and Specificity Article lcsh:Infectious and parasitic diseases 03 medical and health sciences MESH: Zika Virus Infection TaqMan medicine Humans lcsh:RC109-216 MESH: Humans RT-qPCR Outbreak MESH: Chikungunya virus [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology Zika Virus biology.organism_classification Reverse transcriptase MESH: Sensitivity and Specificity Chikungunya Fever [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie Multiplex Polymerase Chain Reaction |
| Zdroj: | International Journal of Infectious Diseases International Journal of Infectious Diseases, Elsevier, 2020, 92, pp.160-170. ⟨10.1016/j.ijid.2019.12.028⟩ International Journal of Infectious Diseases, 2020, 92, pp.160-170. ⟨10.1016/j.ijid.2019.12.028⟩ International Journal of Infectious Diseases, Vol 92, Iss, Pp 160-170 (2020) |
| ISSN: | 1878-3511 1201-9712 |
| Popis: | Highlights • Development and in-house validation of a multiplex RT-qPCR method. • Identification and discrimination of Zika and chikungunya viruses. • Specific and sensitive method applicable to real-life samples. • High coverage of arbovirus strains/isolates. • Testing of the method via a new bioinformatics tool on whole genome sequencing data. Objective The re-emergence and spread of tropical viruses to new areas has raised a wave of concern worldwide. In order to treat patients at an early stage and prevent the diffusion of an outbreak, early diagnosis, and therefore fast and adequate detection, is needed. To this end, a multiplex reverse transcription real-time polymerase chain reaction TaqMan method was designed to detect Zika (ZIKV) and chikungunya (CHIKV) viruses simultaneously. Methods Two methods targeting different genome segments were selected from the literature for each virus. These were adapted for high genome coverage and combined in a four-plex assay that was thoroughly validated in-house. The SCREENED tool was used to evaluate the sequence coverage of the method. Results The full validation approach showed that the new four-plex method allows the specific and sensitive identification and discrimination of ZIKV and CHIKV in routine samples. The combination of two targets per virus allowing almost 100% coverage of about 500 genomes is shown for the first time. Conclusions PCR is a reliable user-friendly technique that can be applied in remote areas. Such multiplex methods enable early and efficient diagnosis, leading to rapid treatment and effective confinement in outbreak cases. They may also serve as an aid for surveillance, and the full validation permits easy method-transfer allowing worldwide harmonization. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|