Mean, standard deviation, range, median and IQR of HC2 RLU and cobas HPV Test Ct for concordant (A) and discordant (B) HC2 and cobas HPV Test results.

<p>Mean, standard deviation, range, median and IQR of HC2 RLU and cobas HPV Test Ct for concordant (A) and discordant (B) HC2 and cobas HPV Test results.</p

Interlaboratory reproducibility between Hospital del Mar (first test) and Hospital de Valme.

<p>Overall test agreement (HR-HPV Positive <i>vs.</i> Negative) <i>kappa value</i> = 0.971.</p

Sensitivity and specificity of HC2 and cobas HPV Test.

<p>HC2 sensitivity for ≥ CIN2: 98.3% (95% CI: 95.1–100); cobas HPV test sensitivity for ≥ CIN2: 98.3% (95% CI: 95.1–100); HC2 specificity for ≥ CIN2: 85.3% (95% CI: 83–87.6); cobas HPV Test specificity for ≥ CIN2: 86.2% (95% CI: 83.9–88.4).</p

Intralaboratory reproducibility.

<p>Overall test agreement (HR-HPV Positive <i>vs.</i> Negative) <i>kappa value = </i>0.957.</p

Mean, standard deviation, range, median and IQR of HC2 RLU and cobas HPV test Ct according to diagnosis, in HPV positive cases.

<p>Mean, standard deviation, range, median and IQR of HC2 RLU and cobas HPV test Ct according to diagnosis, in HPV positive cases.</p

High Resolution Melting Analysis: A Rapid and Accurate Method to Detect <i>CALR</i> Mutations

<div><p>Background</p><p>The recent discovery of <i>CALR</i> mutations in essential thrombocythemia (ET) and primary myelofibrosis (PMF) patients without <i>JAK2/MPL</i> mutations has emerged as a relevant finding for the molecular diagnosis of these myeloproliferative neoplasms (MPN). We tested the feasibility of high-resolution melting (HRM) as a screening method for rapid detection of <i>CALR</i> mutations.</p><p>Methods</p><p><i>CALR</i> was studied in wild-type <i>JAK2/MPL</i> patients including 34 ET, 21 persistent thrombocytosis suggestive of MPN and 98 suspected secondary thrombocytosis. <i>CALR</i> mutation analysis was performed through HRM and Sanger sequencing. We compared clinical features of <i>CALR</i>-mutated <i>versus</i> 45 <i>JAK2/MPL</i>-mutated subjects in ET.</p><p>Results</p><p>Nineteen samples showed distinct HRM patterns from wild-type. Of them, 18 were mutations and one a polymorphism as confirmed by direct sequencing. <i>CALR</i> mutations were present in 44% of ET (15/34), 14% of persistent thrombocytosis suggestive of MPN (3/21) and none of the secondary thrombocytosis (0/98). Of the 18 mutants, 9 were 52 bp deletions, 8 were 5 bp insertions and other was a complex mutation with insertion/deletion. No mutations were found after sequencing analysis of 45 samples displaying wild-type HRM curves. HRM technique was reproducible, no false positive or negative were detected and the limit of detection was of 3%.</p><p>Conclusions</p><p>This study establishes a sensitive, reliable and rapid HRM method to screen for the presence of <i>CALR</i> mutations.</p></div

HRM method.

<p>Examples of melting curves (A) and difference plots (B) obtained for wild-type (WT) and L367fs*46 and K385fs*47 mutations of <i>CALR</i>.</p

Limit of detection of HRM assay.

<p>Melting curves (A) and difference plots (B) of wild-type (blue) and serial dilutions of a <i>CALR</i> K385fs*47 mutant (brown).</p

Immunohistochemistry for CD20 and FOXP3 at study entry and at first response evaluation (1–2 months after finishing treatment).

<p>Gastric biopsies samples from 4 gastric MALT lymphoma patients treated with different schedules. HP: erradication therapy alone; RQ: Rituximab+CHOP; RF: Rituximab+Fludarabine; RB: Rituximab+Bendamustine.</p

Kinetics of CD3+, FOXP3+, ratio FOXP3+/CD3+ and CD20+ cells by immunohistochemistry in responding patients.

<p>Median CD20+ cells were significantly different between cases treated with antibioticis and those treated with fludarabine or bendamustine with or without Rituximab at first response evaluation (p = 0.001) but not later during follow-up.</p