Molecular biology
All qPCR have previously been validated in respiratory samples, based on
the NF-U-47600-2 AFNOR norm.17 Detection of the
following viruses was performed in TW: equine herpesvirus (EHV) -1, −2,
−4 and −5; equine rhinitis virus A (ERAV) and B (ERBV); equine
adenovirus (EAdV) -1 and −2; and equine influenza virus (EIV).
Herpesvirus-2, EHV-5 and ERBV were also assessed in the pooled BALF. Ten
mL of TW and 25 mL of BALF were centrifuged (800 g; 20 min; 4 °C), and
pellets were re-suspended with 1 mL of supernatant. Extraction of viral
nucleic acids and virus specific qPCR were performed using previously
reported assays.18