Analytical and Clinical Performance of a Chikungunya qRT-PCR for Central and South America

Abstract Chikungunya was introduced into the Americas in 2015 causing a pandemic across the continent. Testing during the acute phase of infection relies on qRT-PCR, but available assays have a number of limitations. A qRT-PCR assay specific to the chikungunya E1 gene was designed using sequence dat...

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Published inDiagnostic microbiology and infectious disease Vol. 89; no. 1; pp. 35 - 39
Main Authors Edwards, Thomas, del Carmen Castillo Signor, Leticia, Williams, Christopher, Larcher, Clément, Espinel, Mauricio, Theaker, Jane, Donis, Evelin, Cuevas, Luis E, Adams, Emily R
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.09.2017
Elsevier Biomedical
Subjects
Adolescent
Adult
Aged
Aged, 80 and over
Arboviruses
Chikungunya
Chikungunya Fever - diagnosis
Child
Child, Preschool
Diagnostics
Ecuador
Female
Guatemala
Humans
Infant
Infectious Disease
Internal Medicine
Male
Middle Aged
Molecular diagnostics
Prospective Studies
Real-Time Polymerase Chain Reaction - methods
Retrospective Studies
Reverse Transcriptase Polymerase Chain Reaction - methods
Sensitivity and Specificity
Virology
Young Adult
Guatemala
Ecuador
Chikungunya
molecular diagnostics
diagnostics
arboviruses
Arboviruses
Molecular diagnostics
Diagnostics
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Summary:Abstract Chikungunya was introduced into the Americas in 2015 causing a pandemic across the continent. Testing during the acute phase of infection relies on qRT-PCR, but available assays have a number of limitations. A qRT-PCR assay specific to the chikungunya E1 gene was designed using sequence data from contemporary strains. A probit analysis established the 95% limit of detection as 19.6 copies per reaction. We compared the assay with a US-Centers for Disease Control (CDC) chikungunya qRT-PCR as the reference standard. The assay had a sensitivity and specificity of 98.4% and 100% in 90 samples retrospectively collected in Guatemala. In a further 74 febrile samples prospectively collected in Ecuador and Guatemala the test had a sensitivity and specificity of 100% and 98.4%, respectively. Sequencing the nsp4 gene of the discordant positive sample indicated the presence of chikungunya RNA, and mismatches to the primer binding sites of the CDC assay.
Bibliography:ObjectType-Article-2
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Current address; LGC, Darwin House, Faraday Street, Birchwood Park, Risley, Cheshire, UK.
ISSN:0732-8893
1879-0070
DOI:10.1016/j.diagmicrobio.2017.06.001