RNA-seq of human blood samples from HIV-infected and HIV-uninfected TB contacts
RNA-seq of human blood samples from HIV-infected and HIV-uninfected TB contacts
Three groups of asymptomatic adults at risk of latent tuberculosis infection (household TB contacts, farm workers with occupational exposure to bovine TB and HIV-infected people attending an out-patient clinic) and living in or around Addis Ababa, Ethiopia, were invited to give a blood sample for processing. Principal exclusion criteria were age <18 years; symptoms of active TB; haemoglobin concentration <10 g/dL; and presence of any chest radiograph abnormality consistent with active TB. HIV-infected participants who did not have any contra-indication to IPT were offered a 6-month course of 300 mg isoniazid and pyridoxine 50 mg daily and invited to return to give a second blood sample for the same laboratory tests following treatment completion. 3 mL blood was collected into Tempus Blood RNA Tubes (Thermo Fisher Scientific) to stabilise total RNA for isolation and downstream gene expression analysis, and processed with Tempus Spin RNA Isolation Kits (Thermo Fisher Scientific) to obtain high-quality total RNA. TURBO DNA-free kits (Thermo Fisher Scientific) and GLOBINclear kits (Thermo Fisher Scientific) were then used to remove residual DNA and globin mRNAs, respectively. For genome-wide mRNA sequencing, complementary DNA libraries were generated using Kappa HyperPrep kits (Roche).