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In their Letter to the Editor in this issue, GarciaMontojo, Li and Nath [5] question several technical aspects of our study recently published in Acta Neuropathologica Communications [6]. Our study, in contrast to an earlier report by Nath’s group [11], failed to find elevated expression of HERV-K RNA in postmortem premotor cortex of patients with amyotrophic lateral sclerosis (ALS). Another study by an independent group [12] also failed to confirm elevated expression of HERV-K RNA in postmortem brain tissue of ALS patients. There are a total of five technical issues raised by GarciaMontojo and colleagues in their Letter. Briefly, these are, i) concerns regarding our selection of controls, in particular the number of control subjects with cancer, ii) concerns regarding the number of ALS patients investigated, iii) concerns regarding the RNA integrity numbers (RINs) of the ALS patient tissues and control tissues analysed in our study, iv) concerns regarding our selection of reference genes for RT-qPCR (quantitative real-time reverse transcription polymerase chain reaction) for data normalisation, and v) our use of the RT-qPCR technique rather than alternative methods such as RNASeq (whole transcriptome shotgun sequencing by next-generation sequencing) for analysing the postmortem tissue samples. Each of these questions will be considered in turn:

acta neuropathologica communications

Response to the Letter from Garcia-Montojo and colleagues concerning our paper entitled, Quantitative analysis of human endogenous retrovirus-K transcripts in postmortem premotor cortex fails to confirm elevated expression of HERV-K RNA in amyotrophic lateral sclerosis