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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

  1. a0.0004% Entamoeba dispar, b0.009% Entamoeba histolytica and 0.01% Entamoeba spp., c0.0003% Entamoeba spp., d0.0004% Entamoeba spp.