Diagnosis Of Zika Virus Infection Research Articles

Design, development, and validation of multi-epitope proteins for serological diagnosis of Zika virus infections and discrimination from dengue virus seropositivity.

Zika virus (ZIKV), an arbovirus from the Flaviviridae family, is the causative agent of Zika fever, a mild and frequent oligosymptomatic disease in humans. Nonetheless, on rare occasions, ZIKV infection can be associated with Guillain-Barré Syndrome (GBS), and severe congenital complications, such as microcephaly. The oligosymptomatic disease, however, presents symptoms that are quite similar to those observed in infections caused by other frequent co-circulating arboviruses, including dengue virus (DENV). Moreover, the antigenic similarity between ZIKV and DENV, and even with other members of the Flaviviridae family, complicates serological testing due to the high cross-reactivity of antibodies. Here, we designed, produced in a prokaryotic expression system, and purified three multiepitope proteins (ZIKV-1, ZIKV-2, and ZIKV-3) for differential diagnosis of Zika. The proteins were evaluated as antigens in ELISA tests for the detection of anti-ZIKV IgG using ZIKV- and DENV-positive human sera. The recombinant proteins were able to bind and detect anti-ZIKV antibodies without cross-reactivity with DENV-positive sera and showed no reactivity with Chikungunya virus (CHIKV)- positive sera. ZIKV-1, ZIKV-2, and ZIKV-3 proteins presented 81.6%, 95%, and 66% sensitivity and 97%, 96%, and 84% specificity, respectively. Our results demonstrate the potential of the designed and expressed antigens in the development of specific diagnostic tests for the detection of IgG antibodies against ZIKV, especially in regions with the circulation of multiple arboviruses.

Diagnostic differentiation of Zika and dengue virus exposure by analyzing T cell receptor sequences from peripheral blood of infected HLA-A2 transgenic mice.

Zika virus (ZIKV) is a significant global health threat due to its potential for rapid emergence and association with severe congenital malformations during infection in pregnancy. Despite the urgent need, accurate diagnosis of ZIKV infection is still a major hurdle that must be overcome. Contributing to the inaccuracy of most serologically-based diagnostic assays for ZIKV, is the substantial geographic and antigenic overlap with other flaviviruses, including the four serotypes of dengue virus (DENV). Within this study, we have utilized a novel T cell receptor (TCR) sequencing platform to distinguish between ZIKV and DENV infections. Using high-throughput TCR sequencing of lymphocytes isolated from DENV and ZIKV infected mice, we were able to develop an algorithm which could identify virus-associated TCR sequences uniquely associated with either a prior ZIKV or DENV infection in mice. Using this algorithm, we were then able to separate mice that had been exposed to ZIKV or DENV infection with 97% accuracy. Overall this study serves as a proof-of-principle that T cell receptor sequencing can be used as a diagnostic tool capable of distinguishing between closely related viruses. Our results demonstrate the potential for this innovative platform to be used to accurately diagnose Zika virus infection and potentially the next emerging pathogen(s).

Performance of an Automated Zika IgG Immunoassay in the Detection of Zika IgG Specific Antibodies-A Validation Approach in Samples from Prevalence Areas and Non-Endemic Countries.

The diagnosis of Zika virus infection is complicated and includes testing for nucleic acids and IgM and IgG antibodies, depending on the stage of infection. Zika IgG is an important marker of infection after the acute stage; however, IgG assays can lack specificity due to the similarities between Zika and other flaviviruses. In this study, the diagnostic sensitivity and specificity of the Elecsys® Zika IgG assay were assessed in 496 samples from Zika endemic regions, and specificity only was assessed in 1685 blood screening and diagnostic samples from Zika non-endemic regions. Cross-reactivity was also assessed against a panel of 202 potentially cross-reacting samples. The performance of the Elecsys® Zika IgG assay was compared with the anti-Zika virus ELISA IgG. In the samples from the Zika endemic regions, the Elecsys® Zika IgG assay had 92.88% (95% confidence interval 89.42–95.48) sensitivity and 100% specificity and in the samples from Europe the Elecsys® Zika IgG assay specificity was ≥99.62%. The Elecsys® Zika IgG assay was highly specific in samples from both prevalent and non-endemic regions.

Evaluation of ViroTrack Sero Zika IgG/IgM, a New Rapid and Quantitative Zika Serological Diagnostic Assay.

Dengue virus (DENV) and Zika virus (ZIKV) belong to the flavivirus genus and are antigenically closely related. They also share the same mosquito vector and can cause similar symptoms upon infection. However, DENV and ZIKV infections lead to different clinical sequelae and treatments; therefore, clinicians need rapid and accurate diagnostics capable of distinguishing between the two diseases. Methods: We employed the immuno-magnetic assay technology on a microfluidic cartridge (ViroTrack Sero Zika IgG/IgM) for diagnosis of ZIKV infection based on the aggregation of magnetic nanoparticles. We carried out three serological studies including samples from the Dominican Republic, USA, and Nicaragua, aimed at detecting ZIKV-specific IgG and IgM using the ViroTrack Sero Zika IgG/IgM test. Results: The seroconversion results were comparable with ZIKV IgG and IgM reactivity measured by the commercial ZIKV ELISA kit. The sensitivity and specificity for both ZIKV IgG and IgM tested by the ViroTrack Sero Zika IgG/IgM was approximately 98% and 93%, respectively. Conclusion: Serological detection of ZIKV infection by the new ViroTrack Sero Zika IgG/IgM test shows promising performance and limited cross-reactivity with DENV.

Zika virus detection in amniotic fluid and Zika-associated birth defects

Zika virus infection during pregnancy can cause serious birth defects, which include brain and eye abnormalities. The clinical importance of detection of Zika virus RNA in amniotic fluid is unknown. The purpose of this study was to describe patterns of Zika virus RNA testing of amniotic fluid relative to other clinical specimens and to examine the association between Zika virus detection in amniotic fluid and Zika-associated birth defects. Our null hypothesis was that Zika virus detection in amniotic fluid was not associated with Zika-associated birth defects. We conducted a retrospective cohort analysis of women with amniotic fluid specimens submitted to Colombia's National Institute of Health as part of national Zika virus surveillance from January 2016 to January 2017. Specimens (maternal serum, amniotic fluid, cord blood, umbilical cord tissue, and placental tissue) were tested for the presence of Zika virus RNA with the use of a singleplex or multiplex real-time reverse transcriptase-polymerase chain reaction assay. Birth defect information was abstracted from maternal prenatal and infant birth records and reviewed by expert clinicians. Chi-square and Fisher's exact tests were used to compare the frequency of Zika-associated birth defects (defined as brain abnormalities [with or without microcephaly, but excluding neural tube defects and their associated findings] or eye abnormalities) by frequency of detection of Zika virus RNA in amniotic fluid. Our analysis included 128 women with amniotic fluid specimens. Seventy-five women (58%) had prenatally collected amniotic fluid; 42 women (33%) had amniotic fluid collected at delivery, and 11 women (9%) had missing collection dates. Ninety-one women had both amniotic fluid and other clinical specimens submitted for testing, which allowed for comparison across specimen types. Of those 91 women, 68 had evidence of Zika virus infection based on detection of Zika virus RNA in ≥1 specimen. Testing of amniotic fluid that was collected prenatally or at delivery identified 39 of these Zika virus infections (57%; 15 [22%] infections were identified only in amniotic fluid), and 29 infections (43%) were identified in other specimen types and not amniotic fluid. Among women who were included in the analysis, 89 had pregnancy outcome information available, which allowed for the assessment of the presence of Zika-associated birth defects. Zika-associated birth defects were significantly (P<.05) more common among pregnancies with Zika virus RNA detected in amniotic fluid specimens collected prenatally (19/32 specimens; 59%) than for those with no laboratory evidence of Zika virus infection in any specimen (6/23 specimens; 26%), but the proportion was similar in pregnancies with only Zika virus RNA detected in specimens other than amniotic fluid (10/23 specimens; 43%). Although Zika-associated birth defects were more common among women with any Zika virus RNA detected in amniotic fluid specimens (ie, collected prenatally or at delivery; 21/43 specimens; 49%) than those with no laboratory evidence of Zika virus infection (6/23 specimens; 26%), this comparison did not reach statistical significance (P=.07). Testing of amniotic fluid provided additional evidence for maternal diagnosis of Zika virus infection. Zika-associated birth defects were more common among women with Zika virus RNA that was detected in prenatal amniotic fluid specimens than women with no laboratory evidence of Zika virus infection, but similar to women with Zika virus RNA detected in other, nonamniotic fluid specimen types.

Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1.

The Zika virus (ZIKV) is an emerging flavivirus transmitted to humans by Aedes mosquitoes that can potentially cause microcephaly, Guillain–Barré Syndrome, and other birth defects. Effective vaccines for Zika have not yet been developed. There is a necessity to establish an easily deployable, high-throughput, low-cost, and disposable point-of-care (POC) diagnostic platform for ZIKV infections. We report here an automated magnetic actuation platform suitable for a POC microfluidic sandwich enzyme-linked immunosorbent assay (ELISA) using antibody-coated superparamagnetic beads. The smartphone integrated immunoassay is developed for colorimetric detection of ZIKV nonstructural protein 1 (NS1) antigen using disposable chips to accommodate the reactions inside the chip in microliter volumes. An in-house-built magnetic actuator platform automatically moves the magnetic beads through different aqueous phases. The assay requires a total of 9 min to automatically control the post-capture washing, horseradish peroxidase (HRP) conjugated secondary antibody probing, washing again, and, finally, color development. By measuring the saturation intensity of the developed color from the smartphone captured video, the presented assay provides high sensitivity with a detection limit of 62.5 ng/mL in whole plasma. These results advocate a great promise that the platform would be useful for the POC diagnosis of Zika virus infection in patients and can be used in resource-limited settings.

Clinical relevance of Zika symptoms in the context of a Zika Dengue epidemic

BackgroundThe clinical characteristics of the most frequent arbovirosis (Dengue, Zika, Chikungunya) are very similar, which is a diagnostic challenge for clinicians. ObjectiveTo identify the presence of clinical characteristics related to Zika virus infection confirmed by the laboratory in patients during an epidemic of co-infection with Zika and Dengue viruses. MethodCross-sectional descriptive study of patients with clinical - epidemiological suspicion of Zika virus infection, who were admitted in the Hospital “Joaquín Albarrán” (La Habana, Cuba), during June 1 to October 31 of 2017. Demographic and symptoms and signs were recorded. By PCR for Zika virus (in blood or urine) the disease was confirmed. Results1541 patients were studied. The most frequent symptoms and signs were rash (93.8%), pruritus (77.9%), arthralgia (60.0%), headache (50.8%), myalgia (46.1%), fever (34.7%), asthenia (31.7%), and conjunctivitis (27.9%). Zika virus infection was confirmed in 279 patients (18.1%). Greater frequency of arthralgia, asthenia, and diarrhea was demonstrated in Zika confirmed patients. ConclusionMinor clinical relevance was observed in the symptoms or signs of arboviral disease to support the clinical diagnosis of Zika virus infections by clinician during a Zika-Dengue epidemic. The assessment of the temporality of the onset of Zika and Dengue symptoms of these infections is recommended to assist clinicians in the differential diagnosis.

Implications of the clinical gestational diagnosis of ZIKV infection in the manifestation of symptoms of postpartum depression: a case-control study

BackgroundTraumatic events can trigger postpartum depression. Pregnant women in Brazil today are facing an extremely stressful experience. Thus, the objective here was to analyze the prevalence of symptoms of depression in the immediate postpartum period (IPPD) and associate these symptoms with previous stressful, social, psychological, behavioral, obstetrical, clinical, violent and infectious events, particularly exposure to Zika virus (ZIKV) infection during pregnancy.MethodsThis was a case-control study. The sample comprised 213 puerperal women treated in the maternity ward of a public reference hospital in the Araguaia River Valley of Mato Grosso and Goiás, Brazil. Severe IPPD symptoms were confirmed based on the Edinburgh Postnatal Depression Scale (EPDS). A descriptive statistical analysis was carried out using the Epi Info™ version 7.1.5 suite of software tools and the Statistical Package for Social Sciences (SPSS) version 21.0.ResultsA bivariate analysis revealed a significant association between “severe symptoms of IPPD” and the following variables: “clinical diagnosis of ZIKV during pregnancy” (OR = 13.36; 95% CI = 5.34–33.39); “was separated or divorced in the last year” (OR = 3.58; 95% CI = 1.42–8.99); “suffered an accident in the last year” (OR = 3.32, 95% CI = 1.12–9.82); “suffered emotional violence during pregnancy” (OR = 3.80; 95% CI = 1.81–7.99); “suffered physical violence during pregnancy” (OR = 11.86; 95% CI = 2.07–67.82); “fear of her partner” (OR = 17.90; 95% CI = 3.44–92.99); “dengue fever during pregnancy” (OR = 7.85; 95% CI: 1.66–37.05), and “has a family member diagnosed with mental illness” (OR = 2.54; 95% CI = 1.09–5.93). The multivariate analysis confirmed the association of severe PPD symptoms only with the variables of “clinical diagnosis of ZIKV during pregnancy” (OR = 19.82; 95% CI: 5.35–73.39) and “was separated or divorced in the last year” (OR = 3.92; 95% IC = 1.12–13.63).ConclusionsClinically diagnosed ZIKV during pregnancy may be one of the main events associated with severe IPPD symptoms, showing an almost 20-fold higher chance of occurrence than other factors.

Performance of 2 Commercial Serologic Tests for Diagnosing Zika Virus Infection.

Reliable serologic tests are needed for diagnosis and surveillance of Zika virus infection. We evaluated the Euroimmun and Dia.Pro serologic tests for detection of Zika virus IgM and IgG by using a panel of 199 samples from a region endemic for flaviviruses. Kinetics of Zika virus antibodies were monitored from 300 sequential specimens sampled over a period of 10 months after infection. We observed suboptimal performance; sensitivity for Zika virus IgM was low, especially in the Euroimmun assay (49%), whereas IgM could be detected for months with the Dia.pro assay. The specificity of the Zika virus IgG assays was also low, especially that of Dia.Pro (62%); findings were strongly influenced by the epidemiologic context. These results highlight the complexity of serologic diagnosis of Zika virus infection in regions endemic for flaviviruses. Accurate analysis of the performance of assays is required to adapt and interpret algorithms.

Postnatally Acquired Zika Virus Disease Among Children, United States, 2016-2017.

The clinical findings among children with postnatally acquired Zika virus disease are not well characterized. We describe and compare clinical signs and symptoms for children aged <18 years. Zika virus disease cases were included if they met the national surveillance case definition, had illness onset in 2016 or 2017, resided in a participating state, and were reported to the Centers for Disease Control and Prevention. Pediatric cases were aged <18 years; congenital and perinatal infections were excluded. Pediatric cases were matched to adult cases (18‒49 years). Clinical information was compared between younger and older pediatric cases and between children and adults. A total of 141 pediatric Zika virus disease cases were identified; none experienced neurologic disease. Overall, 28 (20%) were treated in an emergency department, 1 (<1%) was hospitalized; none died. Of the 4 primary clinical signs and symptoms associated with Zika virus disease, 133 (94%) children had rash, 104 (74%) fever, 67 (48%) arthralgia, and 51 (36%) conjunctivitis. Fever, arthralgia, and myalgia were more common in older children (12‒17 years) than younger children (1‒11 years). Arthralgia, arthritis, edema, and myalgia were more common in adults compared to children. This report supports previous findings that Zika virus disease is generally mild in children. The most common symptoms are similar to other childhood infections, and clinical findings and outcomes are similar to those in adults. Healthcare providers should consider a diagnosis of Zika virus infection in children with fever, rash, arthralgia, or conjunctivitis, who reside in or have traveled to an area where Zika virus transmission is occurring.

Validation of serological and molecular methods for diagnosis of zika virus infections

The laboratory confirmation of Zika virus (ZIKV) infection, and the differential diagnosis from other flavivirus infections such as dengue virus (DENV), often requires the use of several diagnostic test types. Cross-reactions and secondary infections complicate the serological diagnosis and specific viral RNA detection assays are often needed for confirming the diagnosis.The aim of this study was to validate serological and molecular methods for diagnosing ZIKV infection. This included the evaluation of a ZIKV RT-qPCR assay for diagnostics that was previously set up for research use and to compare the ZIKV, DENV and TBEV EIA methods. External and in-house controls and pre-characterized sample panels were tested, and also automated and manual nucleic acid extraction methods were compared.A total of ten Finnish traveler patients were diagnosed with acute ZIKV infection during 2015–2017 including one suspected dual DENV and ZIKV infection. These samples along with panels of DENV and tick-borne encephalitis virus (TBEV) infections were used to test the cross-reactive properties of ZIKV, DENV and TBEV IgM assays. Additionally, the diagnosed acute ZIKV patient samples were tested using commercially available diagnostic DENV NS1 antigen assay and a ZIKV NS1 antigen assay intended for research use.The ZIKV RT-qPCR assay was demonstrated to be both specific and sensitive (one genome per reaction) and suitable for routine diagnostic use utilizing automated nucleic acid extraction. Of the tested IgM tests the NS1 antigen-based ZIKV IgM (Euroimmun) assay performed with least cross-reactivity with a specificity of 97.4%. The DENV IgM assay (Focus Diagnostics) had specificity of only 86.1%. The results are in line with previous studies and additionally highlight that also acute TBEV patients may give a false positive test result in DENV and ZIKV IgM assays.

Assay Challenges for Emerging Infectious Diseases: The Zika Experience.

From the perspective of vaccine development, it is imperative to accurately diagnose target infections in order to exclude subjects with prior exposure from evaluations of vaccine effectiveness, to track incident infection during the course of a clinical trial and to differentiate immune reactions due to natural infections from responses that are vaccine related. When vaccine development is accelerated to a rapid pace in response to emerging infectious disease threats, the challenges to develop such diagnostic tools is even greater. This was observed through the recent expansion of Zika virus infections into the Western Hemisphere in 2014–2017. When initial Zika vaccine clinical trials were being designed and launched in response to the outbreak, there were no standardized sets of viral and immunological assays, and no approved diagnostic tests for Zika virus infection. The diagnosis of Zika virus infection is still an area of active research and development on many fronts. Here we review emerging infectious disease vaccine clinical assay development and trial execution with a special focus on the state of Zika virus clinical assays and diagnostics.

Diagnostic performance of commercial IgM and IgG enzyme-linked immunoassays (ELISAs) for diagnosis of Zika virus infection

BackgroundSerologic detection of Zika virus (ZIKV) infections is challenging because of antigenic similarities among flaviviruses.ObjectiveTo evaluate the sensitivity and specificity of commercial ZIKV IgM and IgG enzyme-linked immunoassay (ELISA) kits.MethodsWe used sera from febrile patients with RT-PCR-confirmed ZIKV infection to determine sensitivity and sera from RT-PCR-confirmed dengue cases and blood donors, both of which were collected before ZIKV epidemics in Brazil (2009–2011 and 2013, respectively) to determine specificity.ResultsThe ZIKV IgM-ELISA positivity among RT-PCR ZIKV confirmed cases was 0.0% (0/14) and 12.5% (1/8) for acute- and convalescent-phase sera, respectively, while its specificity was 100.0% (58/58) and 98.3% (58/59) for acute- and convalescent-phase sera of dengue patients, and 100.0% (23/23) for blood donors. The ZIKV IgG-ELISA sensitivity was 100.0% (6/6) on convalescent-phase sera from RT-PCR confirmed ZIKV patients, while its specificity was 27.3% (15/55) on convalescent-phase sera from dengue patients and 45.0% (9/20) on blood donors’ sera. The ZIKV IgG-ELISA specificity among dengue confirmed cases was much greater among patients with primary dengue (92.3%; 12/13), compared to secondary dengue (7.1%; 3/42).ConclusionsIn a setting of endemic dengue transmission, the ZIKV IgM-ELISA had high specificity, but poor sensitivity. In contrast, the ZIKV IgG-ELISA showed low specificity, particularly for patients previously exposed to dengue infections. This suggests that this ZIKV IgM-ELISA is not useful in confirming a diagnosis of ZIKV infection in suspected patients, whereas the IgG-ELISA is more suitable for ZIKV diagnosis among travelers, who reside in areas free of flavivirus transmission, rather than for serosurveys in dengue-endemic areas.

Diagnosis of Zika Virus Infection by Peptide Array and Enzyme-Linked Immunosorbent Assay.

ABSTRACTZika virus (ZIKV) is implicated in fetal stillbirth, microcephaly, intracranial calcifications, and ocular anomalies following vertical transmission from infected mothers. In adults, infection may trigger autoimmune inflammatory polyneuropathy. Transmission most commonly follows the bite of infected Aedes mosquitoes but may also occur through sexual intercourse or receipt of blood products. Definitive diagnosis through detection of viral RNA is possible in serum or plasma within 10 days of disease onset, in whole blood within 3 weeks of onset, and in semen for up to 3 months. Serological diagnosis is nonetheless critical because few patients have access to molecular diagnostics during the acute phase of infection and infection may be associated with only mild or inapparent disease that does not prompt molecular testing. Serological diagnosis is confounded by cross-reactivity of immune sera with other flaviviruses endemic in the areas where ZIKV has recently emerged. Accordingly, we built a high-density microarray comprising nonredundant 12-mer peptides that tile, with one-residue overlap, the proteomes of Zika, dengue, yellow fever, West Nile, Ilheus, Oropouche, and chikungunya viruses. Serological analysis enabled discovery of a ZIKV NS2B 20-residue peptide that had high sensitivity (96.0%) and specificity (95.9%) versus natural infection with or vaccination against dengue, chikungunya, yellow fever, West Nile, tick-borne encephalitis, or Japanese encephalitis virus in a microarray assay and an enzyme-linked immunosorbent assay (ELISA) of early-convalescent-phase sera (2 to 3 weeks after onset of symptomatic infection).

Transverse myelitis associated with Zika virus infection: a Case Report

Title: Transverse myelitis associated with Zika virus infection: a Case Report.: The zika virus belongs to the family flaviviridae, and is transmitted by the arthropod Aedes aegypt. Its major importance is related to the several debilitating neurological manifestations associated with it, such as transverse myelitis.&#x0D; Case: The authors report a case of transverse myelitis in a patient with a previous diagnosis of Zika virus infection. After the image exams and serology, the diagnosis was confirmed and treatment with prednisone was performed with an unsatisfactory clinical outcome.&#x0D; Conclusion: Brazil is the country of greatest concern worldwide due to the thousands of Zika cases with associated neurological complications, causing sequels and physical disabilities. However, cases worldwide have increased substantially.

Zika virus outbreak in Suriname, a report based on laboratory surveillance data.

Introduction : Since the identification of ZIKV in Brazil in May 2015, the virus has spread extensively throughout the Americas. Cases of ZIKV infection have been reported in Suriname since October 2, 2015. Methods : A laboratory-based surveillance system was quickly implemented according to previous experience with the emergence of chikungunya. General practitioners and public health centers located in different districts of Suriname were asked to send blood samples from suspicious cases to Academic Hospital for molecular diagnosis of Zika virus infection. We investigated Zika-related laboratory data collected during surveillance and response activities to provide the first outbreak report in Suriname in terms of time, location and person. Results : A total of 791 molecularly confirmed cases were reported during a 48-week interval from October 2015 to August 2016. The majority of ZIKV-positive cases involved women between 20 and 39 years of age, reflecting concern about Zika infection during pregnancy. The outbreak peaked in mid-January and gradually spread from the district of Paramaribo to western coastal areas. Discussion : This report provides a simple and comprehensive description of the outbreak in Suriname and demonstrates the utility of laboratory data to highlight the spatiotemporal dynamics of the outbreak in that country.

Diagnosis of Zika Virus Infections: Challenges and Opportunities.

Accurate diagnosis of Zika virus (ZIKV) infections has become a pressing need for the effective prevention and control of the epidemic. The findings that ZIKV infections are associated with birth defects and neurologic disease, and that the virus can be sexually transmitted, accentuate the need for accurate diagnostic testing for different applications new to the arbovirus field. Antibody response to related flaviviruses has long been known to be cross-reactive, and antibody detection of ZIKV is nonspecific in populations previously exposed to any of the four dengue viruses or West Nile virus, or vaccinated against yellow fever virus. Therefore, the diagnosis of ZIKV infections has increasingly depended on detection by nucleic acid tests. During the recent epidemic, tests authorized for emergency use have been utilized by public health laboratories and the commercial sector, but a more dependable and responsive diagnostic testing has yet to be developed.

Aptamer-Based ELISA Assay for Highly Specific and Sensitive Detection of Zika NS1 Protein.

We report here a few Zika NS1-binding ssDNA aptamers selected using the conventional SELEX protocol, and their application in an ELISA assay for sensitive diagnosis of Zika NS1 protein. Among the aptamers identified, aptamers 2 and 10 could recognize different binding epitopes of Zika NS1 protein. This complementary in binding site, when coupled with an extraordinarily high binding affinity by 2 (41-nt, KD = 45 pM) and high specificity by 10, was used successfully to construct an ELISA-based assay where 2 and 10 serve as the capture and detection agents, respectively, giving rise to a highly specific detection of Zika NS1 with a detection limit of 100 ng/mL in buffer. Further testing of a few in-house anti-Zika NS1 antibodies show that 2 could also pair with an anti-Zika NS1 antibody. Such aptamer-antibody pairing not only lowers the detection sensitivity by 3 orders of magnitude to 0.1 ng/mL in buffer but also enable highly sensitive detection of as low as 1 and 10 ng/mL of Zika NS1 to be carried out in 10% and 100% human serum, respectively. These results suggest that the selected aptamers would be useful for medical diagnosis of Zika virus infection in various aptamer-based diagnostic devices including ELISA assay.

First imported case of Zika virus infection into Jiangsu Province, China, returning from Ecuador

We report a case of laboratory-confirmed Zika virus (ZIKV) infection imported into China from Ecuador. This is the first documented imported case with ZIKV infection into Jiangsu, and the first imported case into China from Ecuador nationwide. The patient developed fever, rash, conjunctivitis and joint pain after returning. However, due to the less specific clinical symptoms and lack of quick diagnostic methods in elementary healthcare facilities, this imported case was not identified during her first two medical consultations. ZIKV RNA was detected in the urine sample rather than serum sample of the case, and anti-ZIKV IgM was detected from her serum samples collected 4 and 6 days after her onset of symptom. Case isolation and clearance of mosquito-breeding places were implemented instantly when the case was identified. No more human infections were found during a follow-up of 14 days since the case’s isolation. This incident highlights the important role of urine sample in the diagnosis of ZIKV infection and the urgent need for efficient rapid diagnostic method for clinical healthcare providers.

Zika virus-induced neurological critical illness in Latin America: Severe Guillain-Barre Syndrome and encephalitis

Zika virus (ZIKAV) is classically described as causing minor symptoms in adult patients, however neurologic complications have been recognized. The recent outbreak in Central and South America has resulted in serious illness in some adult patients. We report adult patients in Latin America diagnosed with ZIKAV infection admitted to Intensive Care Units (ICUs). MethodsMulticenter, prospective case series of adult patients with laboratory diagnosis of ZIKAV in 16 ICUs in 8 countries. ResultsBetween December 1st 2015 and April 2nd 2016, 16 ICUs in 8 countries enrolled 49 critically ill patients with diagnosis of ZIKAV infection. We included 10 critically ill patients with ZIKAV infection, as diagnosed with RT-PCR, admitted to the ICU. Neurologic manifestations concordant with Guillain-Barre Syndrome (GBS) were present in all patients, although 2 evolved into an encephalitis-like picture. 2 cases died, one due to encephalitis, the other septic shock. ConclusionsDiffering from what was usually reported, ZIKAV infection can result in life-threatening neurologic illness in adults, including GBS and encephalitis. Collaborative reporting to identify severe illness from an emerging pathogen can provide valuable insights into disease epidemiology and clinical presentation, and inform public health authorities about acute care priorities.