Zika virus infections - PDF Document

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  1. Zika virus infections EUROIMMUN test systems for the diagnosis of Zika virus infections

  2. 2 Zika virus Zika virus (ZIKV) belongs to the family of fl aviviruses which encompasses more than 50 different viruses. Dengue, yel- low fever and West Nile viruses are the most prominent fl aviviruses. Zika virus was fi rst isolated in 1947, but only became more known due to a series of epidemics in recent years. symptoms, which usually encompass rash, fever, head- ache, pains in the joints, and conjunctivitis. These symp- toms usually occur 3 – 12 days after the mosquito bite and persist for 2 – 7 days. The disease course is usually mild and self-limiting. The symptoms are very similar to those of dengue or chikungunya virus infections. Since 2013, an increasing number of Zika virus outbreaks in different regions has been registered, for example in South- east Asia, Polynesia and the Pacifi c region, some islands in the Caribbean, and in over 30 countries and regions in North, Central and South America. In Brazil and several other countries, a signifi cant increase in neurological disorders such as Guillain-Barré syndrome was registered during the Zika epidemic 2015 / 2016. Moreo- ver, there was an exceptionally high number of babies with microcephaly. The link between Zika virus infection and the occurrence of neurological disorders and foetal malforma- tions is considered as virtually proven. The topic is currently intensively investigated intensively investigated. SYMPTOMS Fever Headache The virus is transmitted to humans by the bite of mos- quitoes of the Aedes genus. Infected female mosquitoes transmit the virus whilst feeding on blood. Another mode of transmission is perinatal transmission, i.e. transmission from an infected mother to the unborn child. Transmission through sexual intercourse and blood transfusions has also been reported from different countries. Skin rash Red eyes Symptoms Muscle/joints pain Fatigue Zika virus infections proceed asymptomatically in most cases (approx. 80 %). Only in 20 % of cases do patients show Ecuador Guyana Jamaica Curaçao Americ. Samoa Tonga Haiti Peru Marshall Isl. Micronesia St. Vincent & the Grenadines St. Maarten Trinidad & Tobago Aruba Bonaire Bahamas Cayman Isl. USA Turks & Caicos Isl. Antigua & Barbuda St. Eustatius & Saba Brit. Virgin Isl. Singapore St. Kitts & Nevis Montserrat Palau Bolivia Virgin Islands Dom. Rep. Costa Rica Guadeloupe St. Martin Nicaragua Barbados Maledives Samoa Fr. Polynesia New Caledonia Easter Island Cook Islands Malaysia Philippines Cambodia Indonesia Thailand Bangladesh Cuba Fiji St. Barthelemy Belize St. Lucia Argentina Grenada Guinea-Bissau Anguilla Brazil Vanuatu Fiji Colombia Cabo Verde Samoa Solomon Isl. El Salvador Guatemala Mexico Paraguay Suriname Venezuela Fr. Guiana Honduras Martinique Panama Puerto Rico Yap (Micro- nesia) Gabon 2007 - 2009 2012 - 2014 Jan - Oct 2015 Nov 2015 Dec 2015 Jan 2016 Feb 2016 Mar - Jun 2016 Jul - Dec 2016 Spread of Zika virus from 2007 to 2016 (reference: WHO) S d f Zik i f 2007 t 2016 ( f WHO)

  3. 3 Diagnosis Diagnosis of Zika virus infections can be made by direct detection of viral RNA or by indirect detection, i.e. deter- mination of antibodies. Direct detection of the virus in the blood of the patient is possible for maximum one week after onset of symptoms. Specific antibodies are detect- able for a longer period and can indicate both past and acute infections. with other flaviviruses the cross reactivity is only mini- mal. In cases of prior infection or vaccination to another flavivirus, however, a significant cross reactivity must be expected. This can be virtually excluded by the use of highly specific NS1 antigen. Serology Zika virus direct detection (anti-Zika virus antibodies) From approximately day 5 onwards, IgA and / or IgM antibodies can be detected in the serum of infected patients. Approximately 2 – 3 days after the appearance of IgA and / or IgM antibodies, also IgG antibodies are detectable. However, all antibody classes may also appear at the same time. Antibodies can be detected by means of serological tests such as ELISA or indirect immunofluorescence (IIFT). In the interpretation of sero- logical results, the structural similarity of flaviviruses must be taken into account, as this can cause cross reactions of the specific antibodies. In patients without past contact Day 0 Fever onset IgA / IgM Viraemia IgG Viral RNA Course of infection Time window for reliable diagnosis of Zika virus infections The most suitable method for the detection of Zika virus infection depends on the disease stage. In the early phase, viral RNA can be determined. The Zika virus can be detected in blood by RT-PCR for up to one week after onset of symptoms. In infected pregnant women, the virus can be detected also several weeks after this in individual cases (according to Driggers et al., (2016)). Virus detection in urine by PCR is possible for a longer period of time than in serum or plasma. Here, positive results may occur up to 2 – 4 weeks after onset of symptoms. If the infection is older than 7 days, serolog- ical testing is recommended. Antibodies can be detected in the blood of the patient from day 5. Virus detection: Serum or plasma Virus detection: Urine Antibody detection: Serum or plasma 40 34 42 46 36 6 0 38 8 44 2 4 32 10 30 12 26 24 20 22 28 18 14 16 Days after onset of symptoms Days after onset of symptoms Virus detection Virus detection Serology (IgA / IgG / IgM) RT-PCR serum RT-PCR urine ELISA or IIFT + + – / + 1 – 7 – – / + + 8 – 27 – – + from 28

  4. 4 Direct pathogen detection by real-time PCR The EURORealTime Zika Virus PCR provides detection of viral RNA in the early phase of a Zika virus infection. This EURO- IMMUN test system includes reverse transcription of the viral RNA into complementary DNA (cDNA), followed by PCR amplification and fluorescence-based real-time detection of defined ZIKV genome sequences The evaluation of results is performed fully automatically using the EURORealTime Analysis Software. EURORealTime Zika Virus PCR 1. RNA isolation 2. Real-time PCR 3. Analysis and reporting Internal control A B + + – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – Full Ful F Ful Fu u Ful l Full F EURO EURO E EURO EURO URO O EURO U U O O O EURO E E y au a a y a a y tomated with h th h th the Real R R l R R l Rea ea a Rea a Time Time T Ti Ti T Time ime m Time ime e e e Ana Software the the the the the the ly l l lysi ysi ysi y s A A A A Ana Ana A A Ana An n A An lysi i i A B C D E F G H Sample Extracted RNA PCR Mixes contain all required components for reverse transcription and real-time PCR Compatible real-time PCR platforms: LightCycler 480 (Roche), Mx3005P System (Agilent Technologies), 7500 (Fast) Real-Time PCR Instrument (Applied Biosystems) Sample material: Urine, serum EURORealTime Analysis Software Standardised and automated raw data analysis for fast and objective result evaluation  Fully automated report production and result docu- mentation including all internal and external controls  Convenient guidance through the entire workfl ow  Automated layout creation for the PCR plates, including all required controls  Supplements existing real-time PCR platforms, fully compatible with different LIS  Method comparison The sensitivity of the EURORealTime Zika Virus PCR was compared to another CE-IVD-certified PCR test was investigated in a coparative study. For this, samples with 3 different concentrations of Zika virus template were analysed in both systems in triplicate determination. Zika virus RNA was detected with both tests in all three or in two out of three runs. Thus, both systems show an identical analytical sensitivity. Conzentration Zika RNA cp / µl template EURORealTime Zika Virus PCR Real-time PCR Zika test manufacturer A In an external comparison study in a Brazilian labora- tory, 29 clinical serum and 26 urine samples were ana- lysed with the EURORealTime Zika Virus PCR and another CE-IVD-certified PCR test. There was a very high agree- ment of the positive and negative results from both test systems of 95.2% and 97.0%, respectively. detected detected detected detected 3 detected detected not detected detected detected detected 1 Real-time PCR Zika Test manufacturer A detected not detected (29 serum samples, 26 urine samples) detected detected positive negative not detected detected 0.5 20 1 positive EURORealTime Zika Virus PCR detected not detected 1 33 negative

  5. 5 Serological differential diagnosis with ELISA test systems In suspected cases of dengue, chikungunya or Zika virus (DENV, CHIKV, ZIKV) infection, EUROIMMUN recommends differ- ential diagnosis following the procedure described in the scheme below. If the blood sample was taken within 7 days after symptom onset, an additional RT-PCR test on serum or urine for direct pathogen detection should be performed. Indication: (1) Symptoms of a DENV, CHIKV, or ZIKV infection ? Fever, rash, joint pains, neurological symptoms1 (2) Stay in an endemic region during or shortly prior to pregnancy2 Parallel detection of antibodies against DENV, CHIKV and ZIKV using ELISA Dengue Virus NS1 ELISA Anti-Dengue Virus Type 1-4 ELISA (IgG / IgM) Anti-Chikungunya Virus ELISA (IgG / IgM) Anti-Zika Virus ELISA (IgA / IgG / IgM) No indication of CHIKV or ZIKV infection3 No indication of DENV infection + + + + Acute DENV infection likely4,6 CHIKV infection ZIKV infection5,6 DENV infection ¹ e.g. loss of mobility, numbness in the limbs, ascending pareses, facial pareses or loss of muscular refl ex as a sign of Guillain- Barré syndrome (GBS). ² Men who have been to endemic regions and whose partner is pregnant should also be examined, since sexual transmission of Zika virus is possible. ³ In clinically-supported suspected cases and in diagnosis during pregnancy, a follow-up sample should be taken within 1 – 2 weeks; If this is also negative, an acute infection is highly unlikely. 4 Cross reactivity with other fl aviviruses cannot be excluded. In secondary infections with other fl aviviruses, the DENV IgG titer can be above the titer of the virus causing the acute infection. 5 False positive results can occur in sera from patients with acute Plasmodium infections. 6 Possible serological constellations and their relevance in fl avivirus infections (e.g. DENV, ZIKV, tick-borne encephalitis virus, yellow fever virus, West Nile virus, etc.): IgG titer increase in follow-up sample after 1 – 2 weeks IgA* IgM IgG Indication of Acute infection without prior contact with fl avivirus (primary infection) – / +** – / + + yes Acute infection after prior contact with fl avivirus (secondary infection) – / +*** – / + + yes Past infection or previous virus contact – – + no *IgA antibody detection can support the diagnosis of acute ZIKV infections; ** IgG antibodies usually occur together with IgA and / or IgM antibodies or shortly after;***In cases of previ- ous contact with other fl aviviruses, the IgM response can occur with a time delay or with reduced intensity.

  6. 6 Study data of ELISA test systems Sensitivity and specifi city Anti-Zika Virus ELISA (IgG / M) 129 samples were investigated with the Anti-Zika Virus ELISA (IgG) and the Anti-Zika Virus ELISA (IgM). 29 sam- ples originated from patients who had tested positive for Zika virus in examinations of the WHOCC. 100 serum sam- ples of healthy pregnant women were used as reference group. Sensitivity of the Anti-Zika Virus ELISA amounted to 97 %, and specificity to 100 % (taking into account both immunoglobulin classes). When considering the immuno- globulin classes separately, the sensitivity was 73 % for the Anti-Zika Virus ELISA (IgG) and 89 % for the Anti-Zika Virus ELISA (IgM). Depending on the disease stage, it is possible that only one Ig class is present. WHOCC, Hamburg / Routine laboratory, Germany* n = 129 positive borderline negative 28 0 0 positive EUROIMMUN Anti-Zika Virus ELISA (IgG and IgM) together 1 0 0 borderline 0 0 100 negative * 29 samples from patients with Zika virus infection: WHO Collaborating Centre for Arbovirus and Haemorrhagic Fever Reference and Research (WHOCC), Hamburg, Germany; 100 samples from healthy pregnant women: Routine laboratory, Germany For the determination of sensitivity and specificity, a se- cond study was performed using 38 samples from patients whose first samples had been classified as positive by Zika Virus RT-PCR (origin: Dominican Republic, Colombia). The serological investigation was done with samples withdrawn > 5 days after onset of symptoms. 33 patients in whom a dengue virus infection could be confirmed in the first samp- le by direct detection were used as a reference. Due to the positive precharacterisation, the follow-up samples were used to determine the specificity (returning travellers, ori- gin: Germany). Samples from 33 European returning travel- lers were investigated in the same way with the Anti-Zika Virus ELISA (IgG) and the Anti-Zika Virus ELISA (IgM). The sensitivity of the Anti-Zika Virus ELISA considering both immunoglobulin classes was 100% at a specificity of 94%. When observing the immunoglobulin classes seperately, the sensitivity of the Anti-Zika Virus ELISA (IgG) amounted to 100% in both panels. The sensitivity of the Anti-Zika Virus ELISA (IgM) was 27 (Columbian panel), or 87% (panel of European returning travellers). The specificity was 97% for each. Confi rmed Zika virus infection (RT-PCR positive at fi rst withdrawal) / Reference panel (confi rmed dengue virus infection) n = 71 positive borderline negative 38 0 2 positive EUROIMMUN Anti-Zika Virus ELISA (IgG and IgM) together 0 0 0 borderline 0 0 31 negative Confi rmed Zika virus infection (RT-PCR positive at fi rst withdrawal) / Reference panel (confi rmed dengue virus infection) n = 66 positive borderline negative 33 0 2 positive EUROIMMUN Anti-Zika Virus ELISA (IgG and IgM) together 0 0 0 borderline 0 0 31 negative To evaluate the specificity of the Anti-Zika Virus ELISAs a study was performed on 72 patient samples that were seropositive for rheumatoid factors and a variety of other autoantibodies (ANA). 22 additional samples originated from patients with acute EBV infection. Of the total of 94 samples, all sera were negative with the Anti-Zika Virus ELISA (IgG), and four sera were positive with the Anti-Zika Virus ELISA (IgM). Anti-Zika Virus ELISA (IgG) positive Anti-Zika Virus ELISA (IgM) positive Possible infl uencing factors n 22 0 % 4.5 % Acute EBV infection 35 0 % 5.7 % Various autoantibodies 37 0 % 2.7 % Rheumatoid factor The high specifi city and low cross reactivity of the EUROIMMUN ELISA was confi rmed in a study of the Uni- versity Clinic Freiburg and the Bernhard-Nocht Institute for Tropical Medicine (Huzly et al., 2016). Serum samples from European patients with fl avivirus infections or vacci- nations, and samples from patients with acute virus infec- tions were investigated. The study yielded a specifi city of 100 % for the Anti-Zika Virus ELISA (IgG) and 98 % for the Anti-Zika Virus ELISA (IgM). Anti-Zika Virus ELISA positive Cohort (Huzly et al.) n (IgG) IgG n (IgM) IgM 21 0 % 38 0 % TBE virus infection 10 0 % 16 0 % Dengue virus infection 15 0 % 15 0 % Yellow fever vaccine 16 0 % – – Hepatitis C virus infection – – 52 5.8 % Polyclonal IgM

  7. 7 Anti-Zika Virus ELISA (IgA) For investigation of the sensitivity and specificity, a study was performed on 38 patients from ZIKV-endemic regions (origin: Dominican Republic, Colombia) and 11 patients from non-ZIKV-endemic regions (returning tra- vellers, origin: Europe), whose first samples had been classified as positive using Zika-Virus RT-PCR. For the serological investigation, samples were used which were withdrawn > 5 days after onset of symptoms. The refe- rence sera were from 33 patients in whom a dengue virus infection was confirmed by direct detection in the first sample. Due to the positive precharacterisation, the follow-up samples were used for the determination of sensitivity and specificity (returning travellers, origin: Germany). The sen- sitivity was 92 %, with a specificity of 97 %. Confi rmed Zika virus infection (RT-PCR positive at fi rst withdrawal) / Reference panel (confi rmed dengue virus infection) n = 82 positive borderline negative 45 0 1 positive EUROIMMUN Anti-Zika Virus ELISA (IgA) 0 0 0 borderline 4 0 32 negative For evaluation of the specificity of the Anti-Zika Virus ELISA (IgG) a further study was performed on 72 patient samples that were seropositive for rheumatoid factors and a variety of other autoantibodies (ANA). 22 addi- tional samples originated from patients with acute EBV infection. All 93 sera showed a negative result using the Anti-Zika Virus ELISA (IgA). Possible infl uencing factors Anti-Zika Virus ELISA (IgA) positive n 22 0 % Acute EBV infection 35 0 % Various autoantibodies 36 0 % Rheumatoid factor Cross reactivity Due to the use of a highly specifi c, recombinant protein as antigen, cross reactions are virtually ruled out in the EUROIMMUN ELISAs. The investigation of sera panels from clinically and serologically characterised patients with high antibody titers of class IgA, IgG and IgM against other fl aviviruses showed a very reduced cross reactivity. The dengue samples used orig- inate from patients with confi rmed dengue infections as determined by positive PCR and / or positive NS1 determination. Note: Double infection or infection with another fl avivirus at an earlier time are possible, particularly in endemic areas. In this case, positive results are not caused by a cross reactivity of the corresponding antibodies. Anti-Zika Virus ELISA positive Antibodies against (in total > 450) n IgA n IgG n IgM Dengue virus Characterised by direct detection methods (RT-PCR and / or NS1 antigen positive) 86 0.0 % 104 0.0 % 103 0,0 % Clinically and serologically characterised 10 10 % (1 sample) 42 2.4 % (1 sample) 42 0,0 % TBE virus After vaccination 15 0.0 % 135 0.0 % 134 0.0 % Clinically and serologically characterised 54 0.0 % 18 0.0 % 18 0.0 % Yellow fever virus After vaccination (neutralisation test positive) 12 0.0 % 12 0.0 % 12 0.0 % Japanese encephalitis virus Clinically and serologically characterised 13 0.0 % 25 4.0 % 25 0.0 % West Nile virus Characterised by direct detection methods (RT-PCR positive) 40 2.5 % (1 sample) 40 0.0 % 40 0.0 % Clinically and serologically characterised (neutralisation test positive) 16 0.0 % 34 0.0 % 34 2.9 % (1 sample)

  8. 8 Primary and secondary immune response in Zika virus infections At the first contact with a virus, IgA and/or IgM antibodies are formed within 3 – 5 days after infection and remain de- tectable in the blood for some weeks. With a time delay, approximately from day 8 on, an increasing amount of IgG antibodies are secreted into the blood (primary immune response, case examples 1 and 2). Case study 1 8 Patient: Returning traveller from Brazil (PCR positive) IgA 7 ? Antibodies are not yet detectable in the blood in the fi rst serum sample, withdrawn at day 4 after onset of symptoms. In this early phase, PCR diagnostics are indispensable. 6 5 Ratio 4 ? In the follow-up sample (day 9), antibodies of class IgA and IgM are present. In the further course, also IgG antibod- ies will be detectable. 3 IgG IgM 2 Cut-off EUROIMMUN Anti-Zika Virus ELISA Days after onset of symptoms 1 IgA (ratio) IgM (ratio) IgG (ratio) 0 0.3 0.2 0.2 4 0 5 10 15 20 25 30 35 7.9 2.4 1.0 9 Days after onset of symptoms 2.4 0.9 3.0 30 Case study 2 Patient: Returning traveller from Colombia ? IgG levels positive after onset of symptoms. Slow titer increase over a longer time period. After approximately 175 days, the titer slowly decreases, but persists above the cut- off. ? IgM levels are fi rst positive, then decrease over the fur- ther course, and are negative after approximately 120 days. 10 ? In the fi rst serum sample after onset of symptoms, IgA antibodies cannot yet be detected. After a fast increase be- tween the fi rst and the second serum sample (day 9 and 14), the IgA titer decreases again. 8 IgM 6 EUROIMMUN Anti-Zika Virus ELISA Days after onset of symptoms IgA Ratio IgA (ratio) IgM (ratio) IgG (ratio) 4 0.0 8.0 2.0 9 IgG 4.4 7.1 2.8 14 0.6 2.3 3.2 65 2 Cut-off 0.4 0.9 3.5 118 0.3 0.6 3.6 175 0 0.2 0.6 2.1 288 0 50 100 150 200 250 300 350 400 0.2 0.4 1.8 307 Days after onset of symptoms 0.2 0.5 1.9 375 Semiquantitative test evaluation by ratio: Negative: Positive: Borderline: Ratio ≥ 1.1 Ratio ≥ 0.8 < 1.1 Ratio < 0.8

  9. 9 In the case of a second contact with the same virus, the IgG titer increases strongly within the first days of infection. Specific IgM antibodies, however, are only formed in reduced, partly not detectable quantities (secondary immune re- sponse, case examples 3 and 4). Within the immune response, antibodies of class IgA are often formed in parallel to the IgM antibodies. The determination of anti-Zika virus IgA antibodies may support diagnosis of an acute infection and contribute essentially to the clarification of ambiguous findings. Case study 3 Patient: 18 years, male, Colombia ? IgG levels positive already on the 3rd day after onset of symptoms (literature: from day 5 – 6). 7 IgG 6 ? IgM levels negative over the complete period of time. 5 ? IgA positive from day 8 – 35 after onset of symptoms. 4 Ratio EUROIMMUN Anti-Zika Virus ELISA 3 IgA symptoms IgA (ratio) IgM (ratio) IgG (ratio) 0.2 0.1 2.0 3 2 2.9 0.5 5.9 15 Cut-off 1 0.7 0.2 5.2 38 IgM 0.6 0.1 5.0 52 0 0.5 0.1 4.9 66 0 20 40 60 80 100 0.6 0.1 5.1 76 Days after onset of symptoms 0.5 0.1 4.7 95 Case study 4 Patient: 51 years, female, Colombia ? IgG levels positive shortly after onset of symptoms (day 6). After further increase, the titer decreases but remains above the cut-off. 6 IgG 5 ? IgM are never detectable. 4 IgA Ratio ? IgA titer is only positive at the 3rd sample withdrawal, three weeks after symptom onset, then decreases, and is negative approximately after day 60. 3 2 Cut-off EUROIMMUN Anti-Zika Virus ELISA Days after onset of symptoms 1 IgA (ratio) IgM (ratio) IgG (ratio) IgM 0.1 0.02 0.6 0 0 0.6 0.01 2.0 6 0 10 20 30 40 50 60 70 3.4 0.03 4.9 24 Days after onset of symptoms 0.6 0.02 3.3 66 Semiquantitative test evaluation by ratio: Negative: Positive: Borderline: Ratio ≥ 1.1 Ratio ≥ 0.8 < 1.1 Ratio < 0.8

  10. 10 Indirect immunofl uorescence Alongside the ELISA test systems, EUROIMMUN offers indirect immunofl uorescence tests (IIFT) for the diagnosis of Zika virus infections. There are three products available: Anti-Zika Virus IIFT 5 4 3 2 1 Anti-Zika Virus IIFT: For the screening of Zika virus antibodies ? 10 7 9 8 6 Arbovirus Fever Mosaic 2: For differential diagnosis of Zika, dengue, and chikungunya virus infections, which cause similar symptoms. ? Substrates: Virus-infected cells: ZIKV * and non-infected cells Arbovirus Profi le 3: Ideally suited for the investigation of cross reactivity within the group of fl aviviruses. ? Arbovirus Fever Mosaic 2 5 4 3 2 1 Cells infected with different arboviruses are used as test substrate. In a positive reaction, mainly the cytoplasmic regions fl uoresce, showing fi ne- to coarse-granular struc- tures (see fi gure below). Some cells also show a net-like fl uorescence pattern with a dense, perinuclear reactivity. 10 7 9 8 6 Substrates: Virus-infected cells: ZIKV, CHIKV, DENV (Types 1-4)* With these fl avivirus mosaics / profi les, several specifi c anti- bodies can be detected simultaneously. They can be used to support clarifi cation of cross reactivities between different fl aviviruses and enable reliable differential diagnosis in the case of similar clinical symptoms. Arbovirus Profi le 3 4 3 5 2 1 3 5 2 1 4 * ZIKV: Zika Virus, DENV: Dengue Virus, CHIKV: Chikungunya Virus, TBEV: Tick-borne encephalitis virus, WNV: West Nile virus, JEV: Japanese encephalitis virus, YFV: Yellow fever virus Substrates: Virus-infected cells: ZIKV, CHIKV, DENV (Types 1-4), TBEV, WNV, JEV, YFV* Cross reactivity Since complete virus particles are used as antigen in the IIFT products (contrary to the ELISA), cross reactivities are to be expected with the antibodies against viruses of the fl avivirus family. In primary fl avivirus infections, the dom- inant antibody titer of the virus which causes the infection can usually be identifi ed by creating a dilution series of the patient sample. In secondary fl avivirus infections, a high cross reactivity of the IgG antibodies must be expected. The fi nal titers are equal or similar on all fl avivirus substrates. In some cases, the determination of IgM antibodies may allow differentiation. Fluorescence pattern: positive reaction: IgG antibodies against Zika virus.

  11. 11 Automation ELISA EUROIMMUN Analyzer I-2P The Anti-Zika Virus ELISAs are suitable for processing on fully-automated analysis instruments. The tests are vali- dated for the Analyzer I and Analyzer I-2P from EUROIMMUN and the DSX device from Dynex. Automated performance using other fully automated, open system analysis devices is possible. However, the combination should be validated by the user. IIFT EUROIMMUN Sprinter XL The IIF tests for the diagnosis of a Zika virus infection can be processed by the automated laboratory devices Sprinter and Sprinter XL from EUROIMMUN. All steps from the dilution and assignment of samples to the incubation and washing of slides are performed fully automatically. Evalu- ation of the incubated slides at the microscopes is possble, supported by the computer-aided immunofl uorescence microscope EUROPattern. Publications 1. Steinhagen K, Probst C, Radzimski C, Schmidt-Chanasit J, Emmerich P, van Esbroeck M, Schinkel J, Grobusch MP, Goorhuis A, Warnecke JM, Lattwein E, Komorowski L, Deerberg A, Saschenbrecker S, Stöcker W, Schlumberger W. Serodiagnosis of Zika virus (ZIKV) infections by a novel NS1-based ELISA devoid of cross-reactivity with dengue virus antibodies: a multicohort study of assay performance, 2015 to 2016. Euro Surveill. 2016;21(50):pii=30426. 2. Huzly D, Hanselmann I, Schmidt-Chanasit J, Panning M. High specificity of a novel Zika virus ELISA in Europeanpatients after exposure to different flaviviruses. Euro Surveill. 2016;21(16):pii = 30203. 3. Granger D, Gomez LJ, Schimek M, Dubbels M, Mosquera JA, Christensen J, Bistodau S, Strain A, Theel ES. Zika virus Antibody Detection: Evaluation of Three Different Serologic Methodologies. Poster CVS 2016 (USA). 4. Borena W, Hofer T, Stiasny K, Aberle SW, Gaber M, von Laer D, Schennach H. No molecular or serological evidence of Zikavirus infection among healthy blood donors living in or travelling to regions where Aedes albopictus circulates. PLoS ONE 2017 12(5): e0178175. 5. Frank C, Cadar D, Schlaphof A, Neddersen N, Günther S, Schmidt-Chansit J, Tappe D. Sexual transmission of Zika virus in Germany, April 2016. Euro Surveill. 2016; 21(23):pii=30252 6. Zanluca C, Dos Santos CN. Zika virus – an overview. Microbes Infect. 2016 May;18(5):295-301. 7. Johansson MA, Mier-Y-Teran-Romero L, Reefhuis J, Gilboa SM, Hills SL. Zika and the Risk of Microcephaly. N Engl J Med. 2016 May 25 8. Musso D, Gubler DJ. Zika Virus. Clin Microbiol Rev. 2016 Jul;29(3):487-524. 9. Zhang FC, Li XF, Deng YO, Tong YG, Qin CF. Excretion of infectious Zika virus in urine. Lancet Infect Dis. 2016 May 2. 10. Driggers RW, Ho CY, Korhonen, et al. Zika virus infection with prolonged maternal viremia and fetal brain abnormalities. N Engl J Med (2016) March. 11. Fourcade C, Mansuya JM, Dutertre MD, Delobel B. Viral load kinetics of Zika virus in plasma, urine and saliva in a couple returning from Martinique, French West Indies. Journal of Clinical Virology, Jun 2016. 12. Calleri G, Burdino E, Bonora S, Raso R. Zika virus infection in two travelers returning from an epidemic area to Italy, 2016: Algorithm for diagnosis and recommendations, Travel Medicine and Infectious Disease (2016).

  12. Product overview Test systems Order number Anti-Zika Virus ELISA (IgA) EI 2668-9601 A Anti-Zika Virus ELISA (IgM) EI 2668-9601 M Anti-Zika Virus ELISA (IgG) EI 2668-9601 G Anti-Zika Virus ELISA (IgAM) EI 2668-9601 Q Anti-Zika Virus IIFT (IgG or IgM) FI 2668-#### G / M Anti-Zika Virus IIFT EUROPattern (IgG or IgM) FR 2668-#### G / M Arbovirus Fever Mosaic 2 (IgG or IgM) FI 2668-####-1 G / M Arbovirus Fever Mosaic 2 EUROPattern (IgG or IgM) FR 2668-####-1 G / M Arbovirus Profi le 3 (IgG or IgM) FI 2668-####-3 G / M EURORealTime Zika Virus PCR MP 2668-#### EUROIMMUN AG · Seekamp 31 · 23560 Lübeck (Germany) · Tel +49 451/5855-0 · Fax 5855-591 · info@euroimmun.de · www.euroimmun.com HI_2668_I_UK_B04, 02 / 2018