Next Generation Sequencing to determine MneHV7 Transcriptome in pigtailed macaque salivary gland tissues

Purpose: The goal of this study is to determine the expression profile of a new macaque HHV-7 homolog in salivary gland tissues from naturally infected pigtailed macaques Methods: Viral mRNA profiles of 9 different pigtailed macaques naturally infected with the HHV7 homolog provisionnally named MneHV7 were generated by deep sequencing using Illumina Highseq. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: . Hierarchical clustering of differentially expressed genes uncovered several as yet uncharacterized genes that may contribute to retinal function. Data analysis with BWA and TopHat workflows revealed a significant overlap yet provided complementary insights in transcriptome profiling. Conclusions: Our study represents the first detailed analysis of viral transcriptomes associated with the macaque HHV-7 homolog generated by RNA-seq technology. Our results show that NGS offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA content within a cell or tissue. Overall design: Examine MneHV7 viral transcriptome in macaque salivary gland with different viral loads determined by qPCR