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Probe size and fixation time for detecting Porcine circovirus–2 (PCV–2) by in situ hybridization in formalin-fixed, paraffin-embedded lymph nodes from experimentally infected pigs were optimized. In situ hybridization using a 169–base pair (bp) probe detected significantly fewer PCV–2–positive cells than when using 8 other larger probes ( P &lt; 0.05). The difference in hybridization intensity between smaller probes (169 and 225 bp) and larger probes (416, 473, 571, 631, 693, and 753 bp) was statistically significant ( P &lt; 0.05). The PCV–2–positive cells were consistently detected in lymph nodes fixed up to 3 days; thereafter, the number of positive cells declined. The PCV–2–positive cells were detected in lymph nodes fixed for up to 730 days. The difference in hybridization intensity between samples fixed for a short term (1 or 3 days) and a longer term (4–730 days) was statistically significant ( P &lt; 0.05). The data demonstrates that the optimal probe size and fixation time for detecting PCV–2 in formalin-fixed, paraffin-embedded lymph nodes is 473 bp and 1–3 days, respectively.

journal of veterinary diagnostic investigation

Optimal Probe Size and Fixation Time for the Detection of Porcine Circovirus–2 DNA by in Situ Hybridization in Formalin-Fixed, Paraffin-Embedded Tissue