One-step RT-PCR for detection of Zika virus.
BACKGROUND
Zika virus (ZIKV) is an emerging mosquito-borne flavivirus circulating in Asia and Africa. Human infection induces an influenza-like syndrome that is associated with retro-orbital pain, oedema, lymphadenopathy, or diarrhea. Diagnosis of Zika fever requires virus isolation and serology, which are time consuming or cross-reactive.


OBJECTIVE
To develop a one-step RT-PCR assay to detect ZIKV in human serum.


METHODS
An assay targeting the envelope protein coding region was designed and evaluated for its specificity, detection limit, repeatability, and capacity to detect ZIKV isolates collected over a 40-year period from various African countries and hosts.


RESULTS
The assay's detection limit and repeatability were respectively 7.7pfu/reaction and 100% in serum and L-15 medium; none of 19 other flaviviruses tested were detected.


CONCLUSIONS
The assay is rapid, sensitive, and specific to detect ZIKV in cell culture or serum, but needs to be validated for diagnosis using clinical samples.
Faye
Oumar
O
Institut Pasteur de Dakar, 36 Avenue Pasteur, BP 220, Dakar, Senegal.
Faye
Ousmane
O
Dupressoir
Anne
A
Weidmann
Manfred
M
Ndiaye
Mady
M
Alpha Sall
Amadou
A
D014644:Genetic Variation; D012367:RNA, Viral / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000235:genetics*; D000071244:Zika Virus / Q000235:genetics / Q000302:isolation & purification*; D000071243:Zika Virus Infection / Q000175:diagnosis*
D012367:RNA, Viral; D014759:Viral Envelope Proteins