Search for viral agents in cerebrospinal fluid in patients with multiple sclerosis using real-time PCR and metagenomics

使用实时 PCR 和宏基因组学寻找多发性硬化症患者脑脊液中的病毒因子

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作者:Karol Perlejewski, Iwona Bukowska-Ośko, Małgorzata Rydzanicz, Tomasz Dzieciątkowski, Beata Zakrzewska-Pniewska, Aleksandra Podlecka-Piętowska, Agata Filipiak, Krzysztof Barć, Kamila Caraballo Cortés, Agnieszka Pawełczyk, Marek Radkowski, Tomasz Laskus

Abstract

Multiple sclerosis (MS) is a chronic, immune-mediated demyelinating disease of the central nervous system of unclear etiology, but there is some evidence that viral infections could be responsible for triggering autoimmune mechanisms against myelin. We searched for viral RNA and DNA in cerebrospinal fluid (CSF) of 34 MS patients and 13 controls using RT-PCR/PCR against common neurotropic viruses. In addition, shotgun DNA- and RNA-based metagenomics were done in 13 MS patients and 4 controls. Specific quantitative real-time RT-PCR/PCR testing revealed the presence of viral nucleic acid in seven (20.59%) MS patients and in one (7.69%) control patient. In MS patients the most frequently detected was human herpesvirus type 6 (HHV-6; 3 cases; 8.82%); followed by Epstein-Barr virus (EBV; 2 cases; 5.88%), varicella zoster virus (VZV; 1 case; 2.94%) and Enterovirus (EV; 1 case; 2.94%). The single identified virus among controls was EBV (7.69%). DNA and RNA metagenomic assays did not identify any known eukaryotic viruses even though three of the analyzed samples were low-level positive by specific quantitative real-time PCR. In conclusion, we detected the presence of Herpesviridae and occasionally Enteroviridae in CSF from patients with MS but their prevalence was not significantly higher than among controls. Metagenomic analysis seems to be less sensitive than real-time RT-PCR/PCR and it did not detect any potential viral pathogens.

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