A study published in the Journal of Translational Medicine finds that viral infections may be the potential triggers for myelic encephalitis (ME)/chronic fatigue syndrome (CFS) since infectious-like symptoms are present in many of the ME/CFS patients.
]. B19V amplification conditions for both cycles were: initial denaturation–6 min, 95 °C; amplification ; final synthesis–7 min, 72 °C.]. Amplification conditions for both cycles were: initial denaturation–5 min, 95 °C; amplification ; final synthesis –10 min, 72 °C. Following nPCR, amplification products were digested with HindIII restriction endonuclease which cleaves HHV-6B 163 bp amplification product into 66 bp and 97 bp fragments, whereas does not cleave HHV-6A.
Viral load was estimated using DNA extracted from peripheral blood by real-time PCR according to the manufacturer’s instructions. HHV-6A/B load was determined with HHV-6 Real-TM Quant and B19V–with Parvovirus B19 Real-TM Quant kit . HHV-7 load was detected using Human Herpes Virus 7 genomes genesig kit and in-house real-time PCR amplifying HHV-7 U90 and PI15 gene sequences based on a previous report by Prusty et al. [].
The determination of cytokine level in blood plasma was performed according to the manufacturer’s protocol. IL-6 level was detected with eBioscience Human IL-6 Platinium ELISA , IL-10 level – using eBioscience Human IL-10 Platinium ELISA , IL-12 level–with eBioscience Human IL-12p70 Platinium ELISA , and TNF-α level–using Biorbyt Human TNFα ELISA kit . Absorbance was measured with microplate reader at 450 nm and concentration calculated.
The statistical analysis was done by GraphPad Prism 7.0 program. Discrete variables were described as numbers and percentage, and difference in frequency of gender, virus-specific antibodies, and virus presence markers between groups was estimated using Chi-square and Fisher´s exact tests as appropriate. Continuous variables were expressed as average ± standard deviation or median .
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