#Oral #fluid: non-invasive alternative for #parvovirus B19 #diagnosis? (J Clin Virol., abstract)

[Source: Journal of Clinical Virology, full page: (LINK). Abstract, edited.]

Journal of Clinical Virology / Available online 19 May 2019 / In Press, Accepted Manuscript

Oral fluid: non-invasive alternative for parvovirus B19 diagnosis?

Rogier Bodewes a, Jeroen Kerkhof a, Jeroen Cremer a, Daphne B. Gijselaar a, Bettie C.G. Voordouw a,b, Irene K. Veldhuijzen c, Maarten Schipper d, Robvan Binnendijk e

{a} Center for Infectious Disease Research, Diagnostics andlaboratorySurveillance (IDS), National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands; {b} Department of Viroscience, ErasmusMC, Rotterdam, The Netherlands; {c} Centre for Infectious Diseases, Epidemiology and Surveillance, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands; {d}
Department of Statistics, Computer Science and Modelling – SIM, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands; {e} Centre for Immunology of Infectious Diseases and Vaccines, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands

Received 3 September 2018, Revised 18 April 2019, Accepted 17 May 2019, Available online 19 May 2019. DOI: https://doi.org/10.1016/j.jcv.2019.05.008

Under a Creative Commons license



  • Parvovirus B19 (B19 V) causes exanthema in children, similar to rubella and measles virus
  • A B19V-qPCR on oral fluid was evaluated as a non-invasive alternative for serum IgM
  • On basis of 116 children with exanthema, sensitivity of the qPCR on oral fluid reached 70%
  • B19V detection in oral fluid is a suitable public health alternative in the diagnosis of fifth disease




Infections with parvovirus B19 (B19 V) have been associated with a wide range of disease manifestations of which erythema infectiosum (fifth disease) in children is most common. Clinical signs following infection of children with B19 V can be similar to measles and rubella. Laboratory detection of B19 V infections is based on detection of B19V-specific IgM antibodies by enzyme immunoassay (IgM-EIA) and/or B19 V DNA by quantitative PCR (qPCR) on blood samples. The need for invasive sampling can be a barrier for public health diagnostics.


To evaluate the use of a dual target B19V-qPCR directed against the NS1 and VP2 of B19 V on oral fluid samples as a non-invasive alternative for laboratory diagnosis of B19 V infections in children below 12 years of age with exanthema.

Study design

Oral fluid and serum samples were collected from 116 children with exanthema. All serum samples were tested by IgM-EIA/IgG-EIA, while all oral fluid and 56 serum samples were tested by B19V-qPCR.


B19V-specific IgM antibodies were detected in 25 of 116 children in the study. B19V DNA was detected in oral fluid in 17 of the 25 children who were IgM positive, as well as two children who were IgM-equivocal or negative. The child with the equivocal IgM had a high quantity of B19 V DNA in oral fluid (7 log IU/ml), compatible with an acute B19V infection. The IgM-negative child was IgG-positive and 4 log IU/ml B19 V DNA was detected in the oral fluid sample, suggesting an acute infection and a falsely negative IgM. Sample size calculations indicated that oral fluid samples for qPCR should be collected from 2-3 children during outbreaks of exanthema to achieve similar sensitivity as IgM-EIA for one child (≥0.9) to confirm or exclude B19 V.


Results indicate that oral fluid samples are a suitable public health alternative for detection of B19 V infections, potentially lowering the barriers for sampling.

Keywords: parvovirus B19 – PCR – serology – exanthema – children – fifth disease

Keywords: Parvovirus B19; Fifth Disease; Diagnostic tests; Serology.



Generation and Selection of a #Panel of Pan- #Filovirus Single-Chain #Antibodies using Cell-Free Ribosome Display (Am J Trop Med Hyg., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Am J Trop Med Hyg. 2019 May 6. doi: 10.4269/ajtmh.18-0658. [Epub ahead of print]

Generation and Selection of a Panel of Pan-Filovirus Single-Chain Antibodies using Cell-Free Ribosome Display.

Kunamneni A1,2, Clarke EC2, Ye C2, Bradfute SB2, Durvasula R2,1.

Author information: 1 Department of Medicine, Loyola University Medical Center, Chicago, Illinois. 2 Department of Internal Medicine, Center for Global Health, University of New Mexico, Albuquerque, New Mexico.



Filoviruses, which include ebolaviruses and marburgvirus, can cause outbreaks of highly lethal hemorrhagic fever. This disease causes significant morbidity and mortality in humans and non-human primates, with human fatality rates reaching 90% during some outbreaks. Currently, there are a lack of licensed vaccines or antivirals for these viruses. Since early symptoms of filovirus infection mimic more common diseases, there is a strong unmet public health and biodefense need for broad-spectrum filovirus rapid diagnostics. We have generated a panel of mouse single-chain Fv-antibodies (scFvs) to filovirus glycoproteins (GPs) using cell-free ribosome display and determined their cross-reactivity profiles to all known filovirus species. Two scFvs (4-2 and 22-1) were able to detect all known Ebolavirus and Marburgvirusspecies. This is the first report on ribosome display scFvs that can detect a broad set of filovirus GPs, which demonstrates the potential for use in diagnostics.

PMID: 31074409 DOI: 10.4269/ajtmh.18-0658

Keywords: Filovirus; Ebola; Marburg; Diagnostic tests.


Differential #human #antibody #repertoires following #Zika #infection and the implications for #serodiagnostics and disease outcome (Nat Commun., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Nat Commun. 2019 Apr 26;10(1):1943. doi: 10.1038/s41467-019-09914-3.

Differential human antibody repertoires following Zika infection and the implications for serodiagnostics and disease outcome.

Ravichandran S1, Hahn M1, Belaunzarán-Zamudio PF2, Ramos-Castañeda J3, Nájera-Cancino G4, Caballero-Sosa S5, Navarro-Fuentes KR6, Ruiz-Palacios G7, Golding H1, Beigel JH8,9, Khurana S10.

Author information: 1 Division of Viral Products, Center for Biologics Evaluation and Research (CBER), FDA, Silver Spring, MD, 20993, USA. 2 Departamento de Infectología, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, 14080, Mexico. 3 Instituto Nacional de Salud Publica, Cuernavaca, 62100, Mexico. 4 Hospital Regional de Alta Especialidad Ciudad Salud, Tapachula, 30830, Chiapas, Mexico. 5 Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado, Tapachula, 30740, Chiapas, Mexico. 6 Instituto Mexicano del Seguro Social, Tapachula, 30700, Chiapas, Mexico. 7 Comisión Coordinadora de los Institutos Nacionales de Salud y Hospitales de Alta Especialidad, Ministry of Health, Mexico City, 14080, Mexico. 8 Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD, 21701, USA. 9 National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, 20852, USA. 10 Division of Viral Products, Center for Biologics Evaluation and Research (CBER), FDA, Silver Spring, MD, 20993, USA. Surender.Khurana@fda.hhs.gov.



Zika virus (ZIKV) outbreak in Americas led to extensive efforts to develop vaccines and ZIKV-specific diagnostics. In the current study, we use whole genome phage display library spanning the entire ZIKV genome (ZIKV-GFPDL) for in-depth immune profiling of IgG and IgM antibody repertoires in serum and urine longitudinal samples from individuals acutely infected with ZIKV. We observe a very diverse IgM immune repertoire encompassing the entire ZIKV polyprotein on day 0 in both serum and urine. ZIKV-specific IgG antibodies increase 10-fold between day 0 and day 7 in serum, but not in urine; these are highly focused on prM/E, NS1 and NS2B. Differential antibody affinity maturation is observed against ZIKV structural E protein compared with nonstructural protein NS1. Serum antibody affinity to ZIKV-E protein inversely correlates with ZIKV disease symptoms. Our study provides insight into unlinked evolution of immune response to ZIKV infection and identified unique targets for ZIKV serodiagnostics.

PMID: 31028263 DOI: 10.1038/s41467-019-09914-3

Keywords: Zika Virus; Serology; Diagnostic tests.


Towards high quality RT #WGS during #outbreaks using #Usutu virus as example (Infect Genet Evol., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Infect Genet Evol. 2019 Apr 20. pii: S1567-1348(19)30056-5. doi: 10.1016/j.meegid.2019.04.015. [Epub ahead of print]

Towards high quality real-time whole genome sequencing during outbreaks using Usutu virus as example.

Oude Munnink BB1, Kik M2, de Bruijn ND3, Kohl R1, van der Linden A1, Reusken CBEM1, Koopmans M4.

Author information: 1 ErasmusMC, Department of Viroscience, WHO Collaborating Centre for Arbovirus and Viral Hemorrhagic Fever Reference and Research, Rotterdam, the Netherlands. 2 Veterinary Pathology Centre, University of Utrecht, the Netherlands. 3 GD Animal Health, Deventer, the Netherlands. 4 ErasmusMC, Department of Viroscience, WHO Collaborating Centre for Arbovirus and Viral Hemorrhagic Fever Reference and Research, Rotterdam, the Netherlands. Electronic address: m.koopmans@erasmusmc.nl.



Recently, protocols for amplicon based whole genome sequencing using Nanopore technology have been described for Ebola virus, Zika virus, yellow fever virus and West Nile virus. However, there is some debate regarding reliability of sequencing using this technology, which is important for applications beyond diagnosis such as linking lineages to outbreaks, tracking transmission pathways and pockets of circulation, or mapping specific markers. To our knowledge, no in depth analyses of the required read coverage to compensate for the error profile in Nanopore sequencing have been described. Here, we describe the validation of a protocol for whole genome sequencing of USUV using Nanopore sequencing by direct comparison to Illumina sequencing. To that point we selected brain tissue samples with high viral loads, typical for birds which died from USUV infection. We conclude that the low-cost MinION Nanopore sequencing platform can be used for characterization and tracking of Usutu virus outbreaks.

Copyright © 2018. Published by Elsevier B.V.

KEYWORDS: Arboviruses; Nanopore; Sequencing; USUV

PMID: 31014969 DOI: 10.1016/j.meegid.2019.04.015

Keywords: Emerging diseases; Infectious Diseases; Diagnostic tests; Usutu virus.


#Diagnostic #delays in #MERS #coronavirus #patients and #health #systems (J Infect Public Health, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

J Infect Public Health. 2019 Apr 18. pii: S1876-0341(19)30135-2. doi: 10.1016/j.jiph.2019.04.002. [Epub ahead of print]

Diagnostic delays in Middle East respiratory syndrome coronavirus patients and health systems.

Ahmed AE1.

Author information: 1 College of Public Health and Health Informatics, King Saud bin Abdulaziz University for Health Sciences, Riyadh, Saudi Arabia; King Abdullah International Medical Research Center, Riyadh, Saudi Arabia; Ministry of National Guard-Health Affairs, Riyadh, Saudi Arabia. Electronic address: ahmeda5@vcu.edu.




Although Middle East respiratory syndrome coronavirus (MERS-CoV) diagnostic delays remain a major challenge in health systems, the source of delays has not been recognized in the literature. The aim of this study is to quantify patient and health-system delays and to identify their associated factors.


The study of 266 patients was based on public source data from the World Health Organization (WHO) (January 2, 2017-May 16, 2018). The diagnostic delays, patient delays, and health-system delays were calculated and modelled using a Poisson regression analysis.


In 266 MERS-CoV patients reported during the study period, the median diagnostic delays, patient delays, and health-system delays were 5 days (interquartile [IQR] range: 3-8 days), 4 days (IQR range: 2-7 days), and 2 days (IQR range: 1-2 days), respectively. Both patient delay (r = 0.894, P = 0.001) and health-system delay (r = 0.163, P = 0.025) were positively correlated with diagnostic delay. Older age was associated with longer health-system delay (adjusted relative ratios (aRR), 1.011; 95% confidence intervals (CI), 1.004-1.017). Diagnostic delay (aRR, 1.137; 95% CI, 1.006-1.285) and health-system delays (aRR, 1.217; 95% CI, 1.003-1.476) were significantly longer in patients who died.


Delays in MERS-CoV diagnosis exist and may be attributable to patient delay and health-system delay as both were significantly correlated with longer diagnosis delay. Early MERS-CoV diagnosis may require more sensitive risk assessment tools to reduce avoidable delays, specifically those related to patients and health system.

Copyright © 2019. Published by Elsevier Ltd.

KEYWORDS: Coronavirus; Diagnostic delay; Health-system delay; MERS-CoV; Patient delay; Saudi Arabia

PMID: 31006635 DOI: 10.1016/j.jiph.2019.04.002

Keywords: MERS-CoV; Diagnostic tests; Saudi Arabia.


#Unreliable usage of a single #influenza virus #IgM #antibody #assay in #ILI: A retrospective study of the 2016–2018 flu epidemic (PLoS One, abstract)

[Source: PLoS One, full page: (LINK). Abstract, edited.]


Unreliable usage of a single influenza virus IgM antibody assay in influenza-like illness: A retrospective study of the 2016–2018 flu epidemic

Yao Yao , Zhao Zhipeng , Song Wenqi, Li Runqing, Zhu Dong, Qin Kun, Zhao Xiuying

Published: April 22, 2019 / DOI: https://doi.org/10.1371/journal.pone.0215514



We retrospectively analyzed serum IgM antibodies (Abs) to influenza viruses from two tertiary hospitals in Beijing from December 2016 to February 2018. Samples from 36,792 patients, aged 0–98 years, were collected and tested. Among the patients, 923 children from two winter flu seasons were assayed with both antigens and IgM Abs to Flu A and Flu B and assigned as paired groups. Another 2,340 adults and 1,978 children with only antigen tested in the 2016 and 2017 winter flu seasons were named as unpaired groups. IgM Abs-positivity rates in children were 0.80% and 36.57% for Flu A and Flu B, respectively, peaking at 4–5 years of age. For adults, the Flu A and Flu B IgM Abs-positivity rates were 10.34% and 21.49%, respectively, peaking at 18–35 years of age. The trend of temporal distribution between the children and the adults was significantly correlated for IgM Abs to Flu B, but not for Flu A. Compared with unpaired groups, the detection rate of Flu A antigen was significantly higher than IgM Abs in children, whereas frequencies of IgM Abs were higher than antigen in adults. Incidence of Flu B antigen was sharply increased in 2017 winter than in the 2016 winter in both children and adults, but no concomitant increase was observed in IgM Abs to Flu B. For paired children groups, incidence of Flu B antigen in the 2017 flu season was significantly higher than that in the 2016 flu season; in contrast, positive rates of IgM Abs in the 2017 flu season were even lower than those in 2016. Considering antigen detection may reflect the Flu A/Flu B epidemic, our results indicate single-assayed IgM Abs were less effective in the diagnosis of acute influenza virus infection, and the use of this assay for epidemiology evaluations was not supported by these findings.


Citation: Yao Y, Zhipeng Z, Wenqi S, Runqing L, Dong Z, Kun Q, et al. (2019) Unreliable usage of a single influenza virus IgM antibody assay in influenza-like illness: A retrospective study of the 2016–2018 flu epidemic. PLoS ONE 14(4): e0215514. https://doi.org/10.1371/journal.pone.0215514

Editor: Dong-Yan Jin, University of Hong Kong, HONG KONG

Received: December 11, 2018; Accepted: April 3, 2019; Published: April 22, 2019

Copyright: © 2019 Yao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All data files are available from the Fighare datebase, https://figshare.com/s/a8342851717c2acfdd00.

Funding: This study was supported by Beijing Municipal Science & Technology Commission Program of China grant no. Z181100001718148 to Dr. Zhao Xiuying and Beijing Tsinghua Changgung Hospital Fund No. 12018C0003 to Zhu Dong. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.

Keywords: Seasonal Influenza; Serology; Diagnostic tests; China.


Whole-Blood #Testing for #Diagnosis of Acute #Zika Virus #Infections in Routine Diagnostic Setting (Emerg Infect Dis., abstract)

[Source: US Centers for Disease Control and Prevention (CDC), Emerging Infectious Diseases Journal, full page: (LINK). Abstract, edited.]

Volume 25, Number 7—July 2019 / Dispatch

Whole-Blood Testing for Diagnosis of Acute Zika Virus Infections in Routine Diagnostic Setting

Jolanda J.C. Voermans, Suzan D. Pas,1, Anne van der Linden, Corine Geurts van Kessel, Marion Koopmans, Annemiek van der Eijk, and Chantal B.E.M. Reusken2

Author affiliations: Erasmus Medical Center, Rotterdam, the Netherlands



We evaluated the benefit of whole blood versus plasma to detect acute Zika virus infections. Comparison of Zika virus quantitative reverse transcription PCR results in single timepoint whole blood–plasma pairs from 227 patients with suspected Zika virus infection resulted in confirmation of 8 additional patients with Zika virus infection.

Keywords: Zika Virus; Diagnostic tests.