Table 2.

Limits of detection for F. culmorum and F. graminearum in duplex PCR with melting curve analysis.

DNA used as template		Species detected
F. graminearum	F. culmorum	F. graminearum	F. culmorum
5 pg	5 pg	+	+
100 pg	100 pg	-	+
100 pg	10 pg	+	+
10 pg	100 pg	-	+
5 pg to 5 ng	-	+	-
-	5 pg to 5 ng	-	+

Positive detection (+) was defined as the presence of a peak in the melting profile which fulfills these conditions:

(i) its melting point identified by thermocycler software lies within 91.0 ± 1.0°C for F. culmorum and 86.5 ± 1.0°C for F. graminearum

(ii) absolute peak area amounts to at least 100 densitometry units

(iii) the relative peak area in relation to the total area under the melting curve amounts to at least 10%.

Identical results were obtained in the presence of an excess (10 ng) of wheat DNA.