194 chapter 2 Table 42 (continued). Baseline characteristics of included microRNA studies malignant histology, ni reference ni, validation set included indeterminate cytology other inclusion and exclusion criteria sample validation set molecular technique tested microRNAs / developed model validation set malignancy rate PTC FVPTC FTC FTC-OV MTC other n.s. Agretti 2012 53 Bethesda III FLUS TN >10 mm; US-FNAC FNAC sample in TRIzol reagent (Invitrogen Life Technologies) quantitative RT-PCR, Taqman miRNA assay decision tree based on expression values of miR146b, -155, -221 28% 15 Keutgen 2012 72 indeterminate TN; in vivo FNAC; indeterminate cytology upon review; histopathological f/u available FNAC sample in RLT lysis buffer, RNA later solution (Qiagen Inc.) or TRIzol (Invitrogen) RT-PCR, Taqman miRNA assay SVM model based on miR-21, -197, -222 and -328 31% 8 8 4 1 1 Shen 2012 30 Bethesda III AUS TN; extra archived FNAC slide available; corresponding histopathological f/u available archived FNAC slides, RNA extracted with Trizol reagent (Invitrogen) quantitative RT-PCR, Taqman miRNA assay Linear discriminant analysis (LDA) classifier based on miR-30d, -146b, -187 and -221 37% 3 2 6 Vriens 2012 125 indeterminate TN; histopathological f/u available FNAC extra passes or remnant of aspirates with enough cellular material, RNA extracted with Trizol reagent (Invitrogen) quantitative RT-PCR, Taqman miRNA assay Expression of miR-100, -125b, -138, -768-3p 30% 37 Labourier 2015 109 Bethesda III or IV TN; no history of cancer or previous radioiodine therapy; FNAC with sufficient nucleic acids for molecular analysis; histological f/u available consecutive FNAC samples with known mutational status and available histologic f/u miRNA gene expression classifier qualitative positive or negative result based 10-microRNA ThyraMIR™ Thyroid miRNA classifier: miR-29b-1-5p, -31-5p, -1381-3p, -139-5p, -146b-5p, -155, -204-5p, -222-3p, -375, -551-3p 32% 10 18 5 2 Rossi 2016 27 TIR3, excluding Hürthle cell cytology TN; liquid-based cytology; no Hürthle cell nodules; histopathological f/u available prospective FNAC, liquidbased cytology, RNA extracted with miRNeasy Mini Kit (Qiagen) quantitative RT-PCR, KAPA SYBR FAST Universal qPCR Kit (KAPA BIOSYSTEMS) Expression of miR-375, other miRs not significant 52% 3 11 TOTAL 416 393915 3 0 1 37 AUS: atypia of undetermined significance. FLUS: follicular lesion of undetermined significance. FTC: follicular thyroid carcinoma. FTC-OV: FTC, oncocytic variant (Hürthle cell carcinoma). FVPTC: papillary thyroid carcinoma, follicular variant. HN: hyperplastic nodule. MTC: medullary thyroid carcinoma. ni: number of indeterminate thyroid nodules included in our analysis. n.s.: not specified. PS: prospective. PTC: papillary thyroid carcinoma. qPCR: quantitative polymerase chain reaction. RT-PCR: reverse transcription PCR. RS: retrospective.
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