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Enteroviruses in blood of patients with type 1 diabetes detected by integrated cell culture and reverse transcription quantitative real-time PCR

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Abstract

Aims

Enteroviruses (EV) have been associated with type 1 diabetes (T1D), but EV RNA detection has been reported in only a small proportion of T1D patients. We studied whether integrated cell culture and reverse transcription real-time PCR could improve EV detection in blood samples from patients with T1D.

Methods

Blood was collected from 13 patients with T1D. The presence of EV RNA in blood was investigated by using real-time RT-PCR. In addition, plasma and white blood cells (WBC) were inoculated to BGM and Vero cell line cultures. Culture supernatants and cells collected on day 7 and day 14 were tested for EV RNA by real-time RT-PCR. Enterovirus identification was performed through sequencing of the VP4/VP2 region.

Results

Enterovirus RNA was detected in blood by using real-time RT-PCR in only one out of 13 patients. The detection of EV RNA in cultures inoculated with clinical samples (plasma and/or WBC) gave positive results in five other patients. The viral loads were low, ranging from 45 to 4420 copies/ng of total RNA. One isolate was successfully identified as coxsackievirus B1.

Conclusions

Integrated cell culture and reverse transcription real-time PCR can improve the detection rate of EV in blood samples of patients with T1D and can be useful to investigate further the relationship between EV and the disease.

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Acknowledgements

This work was supported by Ministère de l’Education Nationale de l’Enseignement Supérieur et de la Recherche, Université Lille 2 (Equipe d’accueil 3610) and Centre Hospitalier Régional et Universitaire de Lille.

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Authors and Affiliations

  1. Laboratoire de Virologie EA3610, Univ Lille, Faculté de Médecine, CHU Lille, 59000, Lille, France

    Enagnon Kazali Alidjinou, Famara Sane, Agathe Baras, Ilham Moumna, Ilka Engelmann & Didier Hober

  2. Service d’Endocrinologie pédiatrique, CHU Lille, 59000, Lille, France

    Christine Lefevre

  3. Service d’Endocrinologie, CHU de Lille, 59000, Lille, France

    Marie-Christine Vantyghem

Authors
  1. Enagnon Kazali Alidjinou
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  2. Famara Sane
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  3. Christine Lefevre
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  4. Agathe Baras
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  5. Ilham Moumna
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  6. Ilka Engelmann
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  7. Marie-Christine Vantyghem
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  8. Didier Hober
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Correspondence to Didier Hober.

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The authors declared that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants included in the study.

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Managed by Antonio Secchi.

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Alidjinou, E.K., Sane, F., Lefevre, C. et al. Enteroviruses in blood of patients with type 1 diabetes detected by integrated cell culture and reverse transcription quantitative real-time PCR. Acta Diabetol 54, 1025–1029 (2017). https://doi.org/10.1007/s00592-017-1041-7

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  • DOI: https://doi.org/10.1007/s00592-017-1041-7

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