Efficient detection of Zika virus RNA in patients’ blood from the 2016 outbreak in Campinas, Brazil

Judice, Carla C.; Tan, Jeslin J. L.; Parise, Pierina Lorencini; Kam, Yiu‐Wing; Milanez, Guilherme Paier; Leite, Juliana Almeida; Caserta, Leonardo Cardia; Arns, Clarice Weis; Resende, Mariângela Ribeiro; Angerami, Rodrigo Nogueira; Amaral, Eliana; Passini, Renato; Freitas, André Ricardo Ribas; Costa, Fábio Trindade Maranhão; Proença-Módena, José Luiz; Ng, Lisa F. P.

doi:10.1038/s41598-018-22159-2

Cited by 20 publications

(18 citation statements)

References 41 publications

(48 reference statements)

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“…In NHP models, ZIKV-associated birth defects (especially neuropathology) appear to be severe and extremely common [8,68], which may not perfectly recapitulate human infection in which the prevalence of CZS is thought to be approximately 5% [69]. While ZIKV genomic RNA copy numbers in PBMC samples in the sheep study were low, this is consistent with the low-level viremia well-documented among human ZIKV patients [54,70,71,72]. Although RT-PCR does not allow for determination of replicating virus, ZIKV RNA copy number per mL detected in PBMC samples from one infected ewe fluctuated dramatically between D1 and D7 PI, implicating viral replication (Figure 3b).…”

Section: Discussionmentioning

confidence: 63%

Experimental Infection of Pregnant Female Sheep with Zika Virus During Early Gestation

Schwarz

Pozor

et al. 2019

Viruses

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Zika virus (ZIKV) is a vertically and sexually transmissible virus resulting in severe congenital malformation. The goal of this study was to develop an ovine model of ZIKV infection. Between 28–35 days gestation (DG), four pregnant animals were infected with two doses of 6 × 106 PFU of ZIKV; four control animals received PBS. Animals were evaluated for 45 days (D) post-infection (PI) and necropsies were performed. Viral RNA was detected in infected ewe peripheral blood mononuclear cells (PBMC) during the first week PI; however, all fluids and tissues were negative upon culture. Anti-ZIKV IgM (1:400) and neutralizing antibodies were detected in all infected animals. Clinical disease, virus, or ZIKV antibodies were not detected in control ewes. After two weeks PI, fetal loss occurred in two infected animals, and at necropsy, three infected animals had placental petechiation and ecchymosis and one had hydramnion. Fetal morphometrics revealed smaller cranial circumference to crown-rump length ratios (p < 0.001) and relative brain weights (p = 0.038) in fetuses of infected animals compared with control fetuses. Immunophenotyping indicated an increase in B cells (p = 0.012) in infected sheep. Additionally, in vitro experiments using both adult and fetal cell lines demonstrated that ovine cells are highly permissive to ZIKV infection. In conclusion, ZIKV infection of pregnant sheep results in a change in fetal growth and gestational outcomes.

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Section: Discussionmentioning

confidence: 63%

Experimental Infection of Pregnant Female Sheep with Zika Virus During Early Gestation

Schwarz

Pozor

et al. 2019

Viruses

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“…Urine and saliva are useful clinical samples for ZIKV detection, given that ZIKV can be found at higher levels in both saliva and urine as compared to serum [26,27]. However, viral RNA levels are still quite low, with urine samples from the 2016 ZIKV outbreak in Brazil containing 3.66-3568.61 copies [28]. CHIKV can also be found in saliva and urine [29,30], but use of these samples did not enhance the detection rate [29].…”

Section: Discussionmentioning

confidence: 99%

Use of Nanotrap particles for the capture and enrichment of Zika, chikungunya and dengue viruses in urine

et al. 2020

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Nanotrap ® (NT) particles are hydrogel microspheres developed for target analyte separation and discovery applications. NT particles consist of cross-linked N-isopropylacrylamide (NIPAm) copolymers that are functionalized with a variety of chemical affinity baits to enable broad-spectrum collection and retention of target proteins, nucleic acids, and pathogens. NT particles have been previously shown to capture and enrich arboviruses including Rift Valley fever and Venezuelan equine encephalitis viruses. Yet, there is still a need to enhance the detection ability for other re-emerging viruses such as Zika (ZIKV), chikungunya (CHIKV), and dengue (DENV) viruses. In this study, we exploited NT particles with different affinity baits, including cibacron blue, acrylic acid, and reactive red 120, to evaluate their capturing and enrichment capability for ZIKV, DENV and CHIKV in human fluids. Our results demonstrate that CN1030, a NT particle conjugated with reactive red 120, can recover between 8-16-fold greater genomic copies of ZIKV, CHIKV and DENV in virus spiked urine samples via RT-qPCR, superior to the other chemical baits. Also, we observed that CN1030 simultaneously enriched ZIKV, CHIKV and DENV in co-infection-based settings and could stabilize ZIKV, but not CHIKV infectivity in saliva spiked samples. CN1030 enriched viral detection at various viral concentrations, with significant enhancement observed at viral titers as low as 100 PFU/mL for ZIKV and 10 PFU/mL for CHIKV. The detection of ZIKV was further enhanced with NT particles by processing of larger volume urine samples. Furthermore, we developed a magnetic NT particle, CN3080, based on the same backbone of CN1030, and demonstrated that CN3080 could also capture and enrich ZIKV and CHIKV in a dose-dependent manner. Finally, in silico docking predictions support that the affinity between reactive red 120 and ZIKV or CHIKV envelope proteins appeared to be greater than acrylic acid. Overall, our data show that NT particles along with reactive red 120 can be utilized as a pre-processing technology for enhancement of detecting febrile-illness causing viruses.

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“…All cases notified to the Malaysian ZIKV clinical surveillance system had both blood and urine samples tested with rRT-PCR for confirmation of ZIKV infection. Although the duration of detectable ZIKV RNA in serum is relatively short, studies had shown ZIKV RNA remain detectable in urine for a longer duration after becoming undetectable in the serum [33, 35, 52, 53]. Because patients sought medical attention while they still had symptoms, it is unlikely that screening of both serum and urine samples would miss out on positive cases of ZIKV infection.…”

Section: Discussionmentioning

confidence: 99%

Zika virus infection in Malaysia: an epidemiological, clinical and virological analysis

et al. 2019

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Background A major outbreak of the Zika virus (ZIKV) has been reported in Brazil in 2015. Since then, it spread further to other countries in the Americas and resulted in declaration of the Public Health Emergency of International Concern (PHEIC) by World Health Organization. In 2016, Singapore reported its first minor ZIKV epidemic. Malaysia shares similar ecological environment as Brazil and Singapore which may also favor ZIKV transmission. However, no ZIKV outbreak has been reported in Malaysia to date. This study aimed to discuss all confirmed ZIKV cases captured under Malaysia ZIKV surveillance system after declaration of the PHEIC; and explore why Malaysia did not suffer a similar ZIKV outbreak as the other two countries. Methods This was an observational study reviewing all confirmed ZIKV cases detected in Malaysia through the ZIKV clinical surveillance and Flavivirus laboratory surveillance between June 2015 and December 2017. All basic demographic characteristics, co-morbidities, clinical, laboratory and outcome data of the confirmed ZIKV cases were collected from the source documents. Results Only eight out of 4043 cases tested positive for ZIKV infection during that period. The median age of infected patients was 48.6 years and majority was Chinese. Two of the subjects were pregnant. The median interval between the onset of disease and the first detection of ZIKV Ribonucleic Acid (RNA) in body fluid was 3 days. Six cases had ZIKV RNA detected in both serum and urine samples. Phylogenetic analysis suggests that isolates from the 7 cases of ZIKV infection came from two clusters, both of which were local circulating strains. Conclusion Despite similar ecological background characteristics, Malaysia was not as affected by the recent ZIKV outbreak compared to Brazil and Singapore. This could be related to pre-existing immunity against ZIKV in this population, which developed after the first introduction of the ZIKV in Malaysia decades ago. A serosurvey to determine the seroprevalence of ZIKV in Malaysia was carried out in 2017. The differences in circulating ZIKV strains could be another reason as to why Malaysia seemed to be protected from an outbreak.

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Efficient detection of Zika virus RNA in patients’ blood from the 2016 outbreak in Campinas, Brazil

Cited by 20 publications

References 41 publications

Experimental Infection of Pregnant Female Sheep with Zika Virus During Early Gestation

Experimental Infection of Pregnant Female Sheep with Zika Virus During Early Gestation

Use of Nanotrap particles for the capture and enrichment of Zika, chikungunya and dengue viruses in urine

Zika virus infection in Malaysia: an epidemiological, clinical and virological analysis

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