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31.08.2017 | Original Article

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

verfasst von: Enagnon Kazali Alidjinou, Famara Sane, Christine Lefevre, Agathe Baras, Ilham Moumna, Ilka Engelmann, Marie-Christine Vantyghem, Didier Hober

Erschienen in: Acta Diabetologica | Ausgabe 11/2017

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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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Titel: Enteroviruses in blood of patients with type 1 diabetes detected by integrated cell culture and reverse transcription quantitative real-time PCR
verfasst von: Enagnon Kazali Alidjinou
Famara Sane
Christine Lefevre
Agathe Baras
Ilham Moumna
Ilka Engelmann
Marie-Christine Vantyghem
Didier Hober
Publikationsdatum: 31.08.2017
Verlag: Springer Milan
Erschienen in: Acta Diabetologica / Ausgabe 11/2017
Print ISSN: 0940-5429
Elektronische ISSN: 1432-5233
DOI: https://doi.org/10.1007/s00592-017-1041-7

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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

Abstract