Table 7 Comparison of the NGS Results in Terms of Percentage of Species-specific Reads of Investigated Entamoeba spp., Entamoeba histolytica, and Entamoeba dispar Per Total Number of Reads in the Sample with the Respective Largest Standard Deviation above the Mean of All Samples in Multiples of the Standard Deviation (SD), Depicted for the Samples from Patients with Invasive Amebiasis. “Positive” = positive Entamoeba histolytica PCR. “Negative” = negative Entamoeba histolytica PCR. “Uncertain” (for “Specific PCR”) = High cycle threshold value > 35 in real-time PCR with associated uncertain interpretation. “Uncertain” (for “Microscopy in neighboring slides”) = Extremely low parasite density with associated uncertain interpretation. “Match” = NGS result matching the diagnosis “invasive amebiasis”. “Partial match” = NGS result matching the diagnosis “invasive mycosis” on genus level only, so it is neither confirmed nor completely rejected, “Mismatch” = NGS results not matching the diagnosis “invasive amebiasis”, “No match” = No evidence of specific sequences above the respective standard deviation (SD). Highest standard deviations (SD) are shown
From: Next-generation sequencing for hypothesis-free genomic detection of invasive tropical infections in poly-microbially contaminated, formalin-fixed, paraffin-embedded tissue samples – a proof-of-principle assessment
Sample
|
Histological diagnosis
|
Specific PCR [7]
|
Microscopy in neighboring slides [7]
|
NGS > 1st SD
|
NGS > 2nd SD
|
NGS > 3rd SD
|
NGS > 4th SD
|
---|
18
|
Invasive amebiasis
|
Positive
|
Many amebae
|
No match
|
No match
|
No match
|
No match
|
19
|
Invasive amebiasis
|
Negative
|
Negative
|
No match
|
No match
|
No match
|
No match
|
20
|
Invasive amebiasis
|
Positive
|
Many amebae
|
No match
|
Mismatcha
|
No match
|
Matchb
|
21
|
Invasive amebiasis
|
Uncertain
|
Few amebae
|
Partial matchc
|
No match
|
No match
|
No match
|
22
|
Invasive amebiasis
|
Negative
|
Negative
|
No match
|
No match
|
No match
|
No match
|
23
|
Invasive amebiasis
|
Negative
|
Uncertain
|
Partial matchd
|
No match
|
No match
|
No match
|
- a0.0004% Entamoeba dispar, b0.009% Entamoeba histolytica and 0.01% Entamoeba spp., c0.0003% Entamoeba spp., d0.0004% Entamoeba spp.