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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%