Nested polymerase chain reaction for detection of Ehrlichia equi genomic DNA in horses and ticks (Ixodes pacificus)

摘要: Abstract A nested polymerase chain reaction for detecting Ehrlichia equi in horses and ticks (Ixodes pacificus) was developed. major second-round PCR product of 928 bp could be readily visualized ethidium bromide-stained agarose minigels. An internal probe used to verify the identity amplified by non-radioactive (digoxigen-based) Southern blotting: additional confirmation provided DNA sequence analysis. dilution study testing sensitivity indicated that derived from ≤7.6 but > 3 infected neurophils sufficient generate a signal. The specificity examined using panel rickettsiae, which only E. closely-related human granulocytotropic ehrlichia produced bands. In an vivo infection study, detected blood buffy-coat cells experimentally-infected horse on days three through 14 post-inoculation. separate six adult I. pacificus as nymphs had been fed experimentally were found PCR-positive equi.