RSV and HBoV1 co-infection causes more severe pathogenic damage to human respiratory epithelium than single infection

To analyze the differences in HAE gene expression between RSV and HBoV1 co-infection and single infection, total RNA of HAE with “RSV single”, “HBoV1 single”, “RSV and HBoV1” infection at 0 (mock-), 2 and 6 d.p.i. was isolated using the Magzol Reagent (Magen, China) according to the manufacturer's protocol. The quantity and integrity of RNA yield was assessed by using the K5500 (Beijing Kaiao, China) and the Agilent 2200 TapeStation (Agilent Technologies, USA) separately. the mRNA was enriched by oligo dT according to instructions of NEB Next® Poly(A) mRNA Magnetic Isolation Module (NEB,USA), and then fragmented to approximately 200 bp. Subsequently, the RNA fragments were subjected to first strand and second strand cDNA synthesis followed by adaptor ligation and enrichment with a low-cycle according to instructions of NEBNext® Ultra™ RNA Library Prep Kit for Illumina. The purified library products were evaluated using the Agilent 2200 TapeStation and Qubit (Thermo Fisher Scientific, USA). The libraries were sequenced by Illumina (Illumina, USA) with paired-end 150bp.