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Table 4 Environmental test

From: Discrimination of three genetically close Aspergillus species by using high resolution melting analysis applied to indoor air as case study

Sample Sample type Species a Amount of DNA/HRM analysis (ng) b Salmon sperm c Cluster d Percentage of confidence (%) d Number of positive detections e
Control
 NTC water water only water only 0 No N/A /  
 NTCsalmon sperm Salmon sperm only Salmon sperm only 10 Yes N/A /  
 Positive control Pure culture A. creber 25 No 3 95.74 ± 7.07 4/4
 Positive control Pure culture A. creber 25 Yes 3 97.32 ± 3.90 4/4
 Positive control Pure culture A. sydowii 25 No 2 97.91 ± 3.26 4/4
 Positive control Pure culture A. sydowii 25 Yes 2 98.89 ± 0.97 4/4
 Positive control Pure culture A. versicolor 25 No 1 99.16 ± 0.49 4/4
 Positive control Pure culture A. versicolor 25 Yes 1 97.99 ± 2.38 4/4
Environmental sample
 Sample 1 Indoor air sample A. versicolor 19.8 No 1 99.15 ± 0.08 6/8
     Yes 1 99.76 ± 0.01 7/8
   P. chrysogenum      
 Sample 2 Indoor air sample A. versicolor 21.7 No 1 98.10 ± 0.88 5/8 $
     Yes 1 98.93 ± 0.45 7/8
   A. glaucus      
   P. chrysogenum      
   yeast (undetermined)      
 Sample 3 Indoor air sample P. chrysogenum 53.3 No N/A   
     Yes N/A   
 Sample 4 Indoor air sample A. versicolor 50 No 1 98.81 ± 1.90 5/8$
     Yes 1 98.58 ± 0.55 8/8
   infertile mycelium      
   P. chrysogenum      
  1. $At least one repetition is considered as negative due to a confidence below the 95 % threshold
  2. aDetermined with classical methods i.e. plate culture and microscopic analysis (determination and counting) in Libert et al. [13]
  3. bAmount of DNA extracted from air samples of 1.5 m3; DNA amount determined with a Nanodrop® 2000; 5 μl of extracted DNA (5 ng/μl) were used in a 20 μl -HRM analysis
  4. cSalmon sperm (salmon sperm) DNA carrier added (10 ng)
  5. dCluster and % of confidence ± standard deviation (SD) defined with the Biorad Precision Melt Analysis software 1.2 (Temse, Belgium)
  6. eA sample is defined as positive for a specific species, if an amplicon is obtained, if the observed Tm corresponds to the Tm defined by Libert et al. [13] for A. versicolor (i.e., 76.5 ± 0.18 °C) and if the sample is classified in the same cluster as the cluster defined for its respective positive control with a confidence >95%