One-step RT-PCR for detection of Zika virus
Oumar Faye1, Ousmane Faye, Anne Dupressoir
1Institut Pasteur de Dakar, 36 Avenue Pasteur, BP 220, Dakar, Senegal.
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.
Study Design:
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.
Insights
A new one-step RT-PCR assay rapidly and accurately detects Zika virus (ZIKV) in human serum. This sensitive and specific method offers a faster diagnostic alternative for ZIKV infections.
Area of Science:
- Virology
- Molecular Biology
- Public Health
Background:
- Zika virus (ZIKV) is an emerging mosquito-borne flavivirus with increasing global health implications.
- Current ZIKV diagnostic methods, including virus isolation and serology, are time-consuming and can be cross-reactive.
- Human ZIKV infection presents with a range of symptoms, necessitating reliable diagnostic tools.
Purpose of the Study:
- To develop a rapid, one-step reverse transcription-polymerase chain reaction (RT-PCR) assay for ZIKV detection.
- To evaluate the assay's performance in detecting ZIKV in human serum.
Main Methods:
- A one-step RT-PCR assay was designed targeting the ZIKV envelope protein coding region.
- The assay's specificity, detection limit, and repeatability were evaluated.
- The assay was tested against ZIKV isolates spanning 40 years from various African countries and hosts, as well as other flaviviruses.
Main Results:
- The developed RT-PCR assay demonstrated a high detection limit of 7.7 pfu/reaction.
- The assay exhibited 100% repeatability in both serum and L-15 medium.
- Crucially, the assay showed no cross-reactivity with 19 other tested flaviviruses, confirming its specificity.
Conclusions:
- The one-step RT-PCR assay is a rapid, sensitive, and specific tool for ZIKV detection in cell culture and serum.
- Further validation of the assay using clinical samples is recommended for routine diagnostic applications.
- This assay holds promise for improving the speed and accuracy of ZIKV diagnosis.
