Evaluation of Altona Diagnostics RealStar Zika Virus Reverse Transcription-PCR Test Kit for Zika Virus PCR Testing

With the emerging Zika virus (ZIKV) epidemic, serologic diagnosis relies on a labor-intensive IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) and confirmation by a plaque reduction neutralization test (PRNT). To streamline serologic testing, several commercial assays have been developed. Our aim was to compare the commercial Euroimmun anti-ZIKV IgM and IgG assays to the reference MAC-ELISA and PRNT currently in use. Serum specimens submitted to Public Health Ontario Laboratory, Canada, were tested for IgM and IgG using the Euroimmun assays and the results were compared with those from MAC-ELISA. The PRNT was performed on positive or equivocal specimens using either MAC-ELISA or Euroimmun assays, MAC-ELISA-inconclusive specimens, and a convenience sample of specimens negative by both assays (cohort 1). Another set of specimens selected on the basis of PRNT results was subsequently tested by the Euroimmun assays (cohort 2). MAC-ELISA was positive, equivocal, negative, and inconclusive in 57/223, 15/223, 147/223, and 4/223 specimens, respectively. Among the 76 specimens that were MAC-ELISA positive, equivocal, or inconclusive, 30 (39.5%) were Euroimmun IgM and/or IgG positive or equivocal. Among the 147 MAC-ELISA-negative specimens, 136 (92.5%) were Euroimmun IgM and IgG negative. The sensitivity of the combined Euroimmun IgM/IgG against the PRNT was 83% (cohort 1) and 92% (cohort 2), whereas the specificity was 81% (cohort 1) and 65% (cohort 2). The combined Euroimmun IgM/IgG showed good specificity (92.5%) but suboptimal sensitivity (39.5%) compared with that of the MAC-ELISA. However, the sensitivity of the combined Euroimmun IgM/IgG against the PRNT was significantly higher (83 to 92%). More studies are needed before commercial assays are implemented for routine ZIKV serologic diagnosis.

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“…Reference PCR and Altona PCR, as well as NS5 gene PCR and sequencing, were performed on serum specimens as previously described (21).…”

Section: Definitionsmentioning

confidence: 99%

Evaluation of Euroimmun Anti-Zika Virus IgM and IgG Enzyme-Linked Immunosorbent Assays for Zika Virus Serologic Testing

et al. 2017

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“…The challenges of molecular assays to detect ZIKV RNA are that a low limit of detection can result in a high proportion of false negative results and testing conditions (samples, assay reagents, and experimental design) can compromise the sensitivity and specificity of detection. Available real-time PCR kits have varying limits of detection from 30 to 1000 copies/mL (38–40). The Trioplex RT-PCR (CDC) assay detects the ZIKV E gene using TaqMan real-time PCR with 100% positive agreement and a limit of detection of 1.93 × 10 4 genome copy equivalents (GCE)/ml.…”

Section: Discussionmentioning

confidence: 99%

Development of an on-chip detection of Zika virus and antibodies simultaneously using array of nanowells

Semenova

Voigt

Donelan

et al. 2018

Preprint

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24Zika virus (ZIKV) infections are an emerging health pandemic of significant medical importance. 25ZIKV appeared recently in the Americas from Africa via the South Pacific. The current outbreak 26 has garnered attention by exhibiting unique characteristics of devastating neurodevelopmental 27 defects in newborns of infected pregnant women. Current guidelines for ZIKV diagnostics 28 developed by the Center of Diseases Control and Prevention (CDC) consist of nucleic acid 29 testing, plaque reduction neutralization test (PRNT), and a serologic test for IgM detection. To 30better accommodate and comply with these guidelines, we developed a simultaneous on-chip 31 detection of ZIKV and anti-ZIKV antibodies using an array of nanowells. Using on-chip 32 microengraving, we were able to detect anti-ZIKV antibodies and their immunoglobulin isotypes. 33In parallel, applying on-chip real-time PCR with epifluorescence microscopy, we were able to 34 quantify ZIKV viral load as low as one copy. To test clinical samples of patients at the post-35 convalescent stage, we analyzed samples from 8 patients. The on-chip nanowells could 36 effectively identify antibodies that reacted against ZIKV envelope protein and their isotypes with 37 high sensitivity and specificity. The small sample requirement with high specificity and 38 sensitivity and combined molecular and serological tests could potentially be very advantageous 39 and beneficial in accurate detection of Zika infection for better disease monitoring and 40 management.41 42

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“…Detection of anti-ZIKV IgM antibodies was performed using the CDC ZIKV MAC-ELISA, an IgM-antibody capture assay, and ZIKV RNA detection in serum was performed using the Altona RealStar ® RT-PCR, as described previously [16]. To test for DENV and chikungunya virus (CHKV) RNA, we adapted the assay previously described by Pongsiri and colleagues [17].…”

Section: Virus and Antibody Detectionmentioning

confidence: 99%

Presence of Flavivirus Antibodies Does Not Lead to a Greater Number of Symptoms in a Small Cohort of Canadian Travelers Infected with Zika Virus

Kozak

Goneau

DeLima

et al. 2020

Viruses

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Zika virus (ZIKV) is a mosquito-borne flavivirus associated with a febrile illness as well as severe complications, including microcephaly and Guillain-Barré Syndrome. Antibody cross-reactivity between flaviviruses has been documented, and in regions where ZIKV is circulating, dengue virus (DENV) is also endemic, leaving the potential that previous exposure to DENV could alter clinical features of ZIKV infection. To investigate this, we performed a retrospective case-control study in which we compared Canadian travellers who had been infected with ZIKV and had serological findings indicating previous DENV or other flavivirus exposure (n = 16) to those without any previous exposure (n = 44). Patient samples were collected between February 2016 and September 2017 and submitted to Public Health Ontario for testing. ZIKV infection was determined using real-time RT-PCR and antibodies against DENV were identified by the plaque-reduction neutralization test. The mean time from symptom onset to sample collection was 5 days for both groups; the magnitude of viremia was not statistically different (Ct values: 35.6 vs. 34.9, p-value = 0.2). Clinical scores were also similar. Our findings indicate that previous DENV or other flavivirus exposure did not result in greater viremia or a higher illness score.

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Evaluation of Altona Diagnostics RealStar Zika Virus Reverse Transcription-PCR Test Kit for Zika Virus PCR Testing

Cited by 29 publications

References 24 publications

Evaluation of Euroimmun Anti-Zika Virus IgM and IgG Enzyme-Linked Immunosorbent Assays for Zika Virus Serologic Testing

Evaluation of Euroimmun Anti-Zika Virus IgM and IgG Enzyme-Linked Immunosorbent Assays for Zika Virus Serologic Testing

Development of an on-chip detection of Zika virus and antibodies simultaneously using array of nanowells

Presence of Flavivirus Antibodies Does Not Lead to a Greater Number of Symptoms in a Small Cohort of Canadian Travelers Infected with Zika Virus

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