Multiple <i>KRAS</i> mutations according to different techniques.

<p>PDAC, Pancreatic Ductal AdenoCarcinoma; IPMN, Intraducatal Pancreatc Mucinous Neoplasia; NA, end-point not available.</p

Example of molecular results in a <i>KRAS</i> exon 2 mutated sample (case #57, Table 4) with discordant results between the three techniques.

<p>A) Electropherogram obtained using Sanger sequencing. The mutation is not identified. B) Using ASLNAqPCR the <i>KRAS</i> G12D mutation is identified by the right curve (G12D). The left curve indicates the wild-type allele (WT). The ratio between the two curves corresponds to ∼6% of mutated alleles. C) Profile obtained using 454-NGS, the <i>KRAS</i> G12D mutation is identified by the vertical green bar. The percentage of mutated alleles is indicated on the left y axis while the total number of reads on the right one.</p

Proposed algorithm for the detection of <i>KRAS</i> mutations in EUS-FNA material from pancreatic lesions.

<p>Ex, Exon; WT, Wild-Type.</p

Percentage of mutated <i>KRAS</i> samples according to preoperative cytology evaluation.

<p>ASLNAqPCR, Allele Specific Locked Nucleic Acid qPCR; NGS, Next Generation Sequencing; PDAC, Pancreatic Ductal AdenoCarcinoma; Not PDAC, malignant neoplasia but not Pancreatic Ductal Adenocarcinoma; NA, cytology not available.</p

Example of molecular results in a <i>KRAS</i> exon 3 mutated sample (case #37, Table 4).

<p>A) Electropherogram obtained using Sanger sequencing. The <i>KRAS</i> Q61H mutation is identified by the smaller pick pointed by the arrow. B) No <i>KRAS</i> mutation was detected using ASLNAqPCR results: only the curve corresponding to the wild-type allele is visible (WT). C) Profile obtained using 454-NGS, the <i>KRAS</i> Q61H mutation is identified by the vertical blue bar. The percentage of mutated alleles is indicated on the left y axis while the total number of reads on the right one.</p

Statistical performance of <i>KRAS</i> molecular analysis using 454-NGS starting only from FNA material or adding the results obtained in DNA extracted from cytologic smears (in bold).

<p>SPEC, Specificity; SENS, Clinical Sensitivity; PPV, Positive Predictive Value; NPV; Negative Predictive Value; ACC; Accuracy; FDR, False Discovery Rate.</p

Example of molecular results in a <i>KRAS</i> exon 2 mutated sample.

<p>A) Electropherogram obtained using Sanger sequencing. The <i>KRAS</i> G12D mutation is identified by the smaller peak pointed by the arrow. B) Using ASLNAqPCR results the <i>KRAS</i> G12D mutation is identified by the right curve (G12D). The left curve indicates the wild-type allele (WT). C) Profile obtained using 454-NGS, the <i>KRAS</i> G12D mutation is identified by the vertical green bar. The percentage of mutated alleles is indicated on the left y axis while the total number of reads on the right one.</p