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Zika Virus Research

 Detection of Zika viral genomic RNA with RNAscope probe #464531 (red punctate dots).  The Vero cells sample was infected with ZIKV, MR766 strain.   Data courtesy of Dr. Noriko Nakajima, National Institute of Infectious Disease, Japan

Detection of Zika viral genomic RNA with RNAscope probe #464531 (red punctate dots). The Vero cells sample was infected with ZIKV, MR766 strain. Data courtesy of Dr. Noriko Nakajima, National Institute of Infectious Disease, Japan

Detection of Zika viral cRNA (- strand) with RNAscope<sup>®</sup> probe #464541 ( red punctate dots).  The Vero cells sample was infected with ZIKV, MR766 strain.  Data courtesy of Dr. Noriko Nakajima, National Institute of Infectious Disease Japan

Detection of Zika viral cRNA (- strand) with RNAscope® probe #464541 ( red punctate dots). The Vero cells sample was infected with ZIKV, MR766 strain. Data courtesy of Dr. Noriko Nakajima, National Institute of Infectious Disease Japan

As a result of the current  Zika virus outbreak, researchers are accelerating research into the biology of the virus and the pathology of human infection with the goals of understanding the causes of the clinical syndrome, developing reliable detection methods, and finding effective treatments.  

Why use RNAscope® assay for zika virus research?

RNAscope® ISH, an innovated ISH assay, offers researchers a robust and accurate method to detect even low levels of viral RNA in virtually any tissue samples.  The ability to detect the virus within the context of the infected tissue will advance the understanding of the zika viral biology. 

Morphologic context—Performed in situ, RNAscope® assay preserves the morphology of the infected tissues, enabling detection of the viral RNA in the morphological context.  Miner et al., 2016 demonstrated this in detection of Zika in eyes of mouse fetuses

Proven for viral detection—Numerous publications citing the use of RNAscope® in situ hybridization in viral studies spanning HPV, EPV, HIV, SIV, HCV, MERS etc.  RNAscope® assay enables virologists to bypass time consuming and often non-yielding antibody development.  

Proven for viral replication detection and localization-- read Bhatnagar J et al 2017

Versatile probe design for low viral count—When working with zika or other viruses of extremely low copy number, ACD can customize the probe design strategy to offset this challenge; for example by designing probes with more than the typical 20 Z probe pairs.

Specificity—The highly specific, proprietary RNAscope® double Z probe design, eliminates cross reactivity with other flavivirus RNA, enabling reliable differentiation from highly homologous viruses such as dengue.

Sensitivity—The ultrahigh sensitivity of RNAscope® assay and ability to detect single RNA molecules makes it ideal for in situ detection of low level tissue infection.  Read Rosenberg et al (2017

Featured Publications

ACD is proud that our customers have used RNAscope Assay and published their research findings. 

 

Date/Journal Title and details

July 2016

Read it in the news

 

New England Journal of Medicine

Miscarriage associated with Zika virus infection

van der Eijk AA et all detected ZIKA viral RNA with the use of virus-specific RNAscope probes that revealed staining in the amniotic epithelium and the fetal mesenchymal cells, particularly in the perichondrium.

SEPT 2016

See the story

Cell Reports

Zika virus infection in mice causes panuveitis with shedding of virus in tears

Miner et al., 2016 detected Zika virus in eyes of mouse fetuses--specifically in the retina, optic nerve and cornea (figure 6)

SEPT 2016

Archives of Path & Lab Med

Placental pathology of Zika virus: viral infection of the placenta induces villous stromal macrophage proliferation and hyperplasia

Rosenberg et al (2017) applied In situ hybridization using a Zika virus RNAscope Probe 463781 demonstrated scattered, strongly positive staining cells within the villous stroma of the chorionic villi (Figure 4), which were presumably Hofbauer cells.

OCT 2016

Read it in the news

Nature Medicine

Zika viral dynamics and shedding in rhesus and cynomolgus macaques

 

OCT 2016

See the story

Nature

Zika virus infection damages the testes in mice

Govero et al (2016) used RNAscope Probe 467871

NOV 2016

Read it in the news

Nature

Neutralizing human antibodies prevent Zika virus replication and fetal disease in mice.

Sapparapu et al (2016) used RNAscope Probe 467771

 

DEC 2016

Read it in the news

Emerging Infectious Disease

Zika virus RNA replication and persistence in brain and placental tissue

Bhatnagar J et al (2017) used RNAscope Probes 468361 and 469541 and demonstrated not only detection of the virus but also detection of viral replication. They isolated the viral replication to neuronal cells in fetal brain tissues.

JAN 2017

 

PLOS

Neuropathogenesis of Zika Virus in a Highly Susceptible Immunocompetent Mouse Model after Antibody Blockade of Type I Interferon

ZIKA Virus Genome  Structure

The zika virus is a sense single-stranded RNA virus (sssRNA)  (also described as SSRNA positive sense virus, no DNA stage)  . Its  >10,000 bases code for three structural proteins (capsid (C), precursor membrane (prM), envelop (E)) and seven nonstructural proteins (NS). The NS2A, NS2B, NS4A, and NS4B proteins are smaller, hydrophobic proteins while NS1, NS3, and NS5 are large and highly conserved.  

ACD can design RNAscope® ISH probes against unique regions of these publically available sequences or proprietary sequences that you may have. We can design it to the sense or antii-sense strand.  Depending on your study, the probe design can target specific gene regions or span across the genome. We will also work with you to design positive and negative controls. 

We can provide full consultation for your Zika research project.

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