A diagnostic polymerase chain reaction assay for Zika virus

Abstract

Zika virus (ZIKV) is a mosquito-borne flavivirus. Infection results in a dengue-like illness with fever, headache, malaise, and a maculopapular rash. Nearly all cases are mild and self-limiting but in 2007, a large outbreak of ZIKV was reported from the island of Yap (in Micronesia, northwest of Indonesia). Singapore is already endemic for dengue, and its impact on public health and economic burden is significant. Other dengue-like infections (e.g., Chikungunya virus) are present. Yet only 10% of reported dengue cases have laboratory confirmation. The identification and control of other dengue-like, mosquito-transmitted infections is thus important for the health of Singapore's population, as well as its economy. Given that ZIKV shares the same Aedes mosquito vector with both dengue and Chikungunya, it is possible that this virus is present in Singapore and causing some of the mild dengue-like illness. A specific and sensitive one-step, reverse transcription polymerase chain reaction (RT-PCR) with an internal control (IC) was designed and tested on 88 archived samples of dengue-negative, Chikungunya-negative sera from patients presenting to our hospital with a dengue-like illness, to determine the presence of ZIKV in Singapore. The assay was specific for detection of ZIKV and displayed a lower limit of detection (LoD) of 140 copies viral RNA/reaction when tested on synthetic RNA standards prepared using pooled negative patient plasma. Of the 88 samples tested, none were positive for ZIKV RNA, however, the vast majority of these were from patients admitted to hospital and further study may be warranted in community-based environments.

Outbreak of Zika Virus Infection, Chiapas State, Mexico, 2015, and First Confirmed Transmission byAedes aegyptiMosquitoes in the Americas

“…All RNA samples were tested using a 1-step reverse transcription-polymerase chain reaction (RT-PCR) with the Qiagen OneStep RT-PCR kit (Qiagen, Valencia, California) and previously described ZIKV-or DENV-specific primers [19]. Quantitative RT-PCR was also performed, using the Taqman RNA-to-C 1-step kit (Applied Biosystems, San Francisco, California) and CHIKV-specific primers [17].…”

Zika Virus

“…Further testing on known Zika virus plasma aliquots is needed. In addition, a wider and larger sample of patients needs to be studied in the community [ 31 ]. In this regard, it should be noted that Zika virus infection could be masked and not easily confi rmed.…”

Zika Virus Infection Acquired During Brief Travel to Indonesia

“…1,3 Diagnosis can be confirmed by demonstration of specific antibodies against Zika virus in serum or by PCR detection of the virus. [4][5][6] Detailed serologic testing of the Yap Islands Zika virus outbreak in 2007 showed limited cross-reactivity with IgM against other flaviviruses, including dengue virus. 7 To our knowledge, this is the first case of Zika virus infection reported in a returned traveler to Australia, although serologic evidence of Zika virus infection has been reported in Java, Indonesia.…”