Fig. 3

Nested PCR detection of Vavraia-like microsporidian DNA from Eurycea salamanders from SMARC. a Agarose gel image of PCR products derived from the frozen tissues of the six salamander specimens collected in 2013 outbreak for initial diagnosis and molecular detection. Negative control (−) used pure water. b Agarose gel image of PCR products derived from a selected number of archived salamander specimens indicating both negative and positive detection. Negative (−) and positive (+) controls used DNA isolated from microsporidian-negative and -positive salamander specimens. Nested PCR reactions used primer pairs Microsp_Salam_S1F and Microsp_Salam_S1R for primary PCR, and Microsp_Salam_S2F and Microsp_Salam_S2R for secondary PCR as described in Additional file 1: Table S1