FIG. 6.

Detection and virus isolation from clinical samples. (A) Phase-contrast and fluorescent photomicrographs of cytopathic effects due to rRPV^KOL-RRegfpR isolated from PBMCs of infected cattle. PBMCs were isolated from cattle infected for 4 days with rRPV^KOL-RRegfpR and overlaid onto B95a cells. Photomicrographs were taken 48 hpi using a digital CCQ camera (Hammatsu) attached to an Eclipse TE300 UV microscope (Nikon). Magnification, ×30. Total RNA was isolated from PBMCs and eye swabs obtained from rRPV^KOL-RRegfpR infected cattle 0, 2, 4, 7, 8 and 9 dpi. PCR products were amplified using morbillivirus universal P and β-actin primers from cDNA prepared from the total RNA. Amplicons were examined by DNA gel electrophoresis. Arrows indicate 500-bp DNA size markers. (B) Amplicons obtained from PBMC samples using the morbillivirus universal P primers (upper panels) and β-actin controls (lower panels). (C) Amplicons obtained from eye swab samples using the morbillivirus universal P primers (upper panels) and β-actin controls (lower panels).