Winter, Alexander and Engels, Svenja and Goos, Philipp and Süykers, Marie-Christin and Henke, Rolf-Peter and Gerullis, Holger and Wawroschek, Friedhelm
(2018)
Detection of CK19 mRNA using one-step nucleic acid amplification (OSNA) in prostate cancer: preliminary results.
Journal of cancer, 9 (24).
pp. 4611-4617.
ISSN 1837-9664
Abstract
Background: Accurate histopathological evaluation of lymph nodes (LNs) is essential for reliable staging
in prostate cancer. In routine practice, conventional techniques only examine parts of the LN. Molecular
nodal staging methods are limited by their high costs and extensive time requirement. One-step nucleic
acid amplification (OSNA) determines the metastatic status of the complete LN and allows for rapid
intraoperative detection of LN metastases. OSNA has been proposed for diagnosis of LN metastases
from breast cancer by quantifying the CK19 mRNA copy number. To provide basic data for OSNA
development for prostate cancer, we conducted an investigation of CK19 and OSNA in prostate cancer
specimens.
Methods: OSNA is based on a short homogenization step and subsequent automated amplification of
CK19 mRNA directly from the sample lysate, with results available in 30–40 min. A total of 20 prostate
cancer specimens from consecutive patients with intermediate or high-risk prostate cancer
(Gleason-Score ≥7) were investigated by both OSNA and conventional histopathology (H&E staining,
CK19 immunohistochemistry). OSNA was performed on frozen samples using a ready-to-use
amplification kit in an automated real-time detection system. Samples were defined as ‘negative’ or
‘positive’ according to mRNA copy number: >5000 copies/μl (++), 250–5000 copies/μl (+), and <250
copies/μl (-).
Results: Histopathological analysis confirmed prostate cancer in all samples: Gleason score 7 (n=11),
Gleason score 8 (n=2), and Gleason score 9 (n=6). Gleason score could not be given for one patient who
previously underwent hormonal treatment. OSNA analysis detected CK19 expression in 100% of the
specimens and high numbers of CK19 mRNA copies in all cases (9 samples ++; 11 samples +).
Immunohistochemistry confirmed CK19 expression in 19 of 20 cases. In the immunohistochemistry
CK19-negative patient, a Gleason score 9 prostate cancer was diagnosed.
Conclusions: This is the first study using OSNA to detect CK19 expression in prostate cancer. Initial
data indicate that this rapid method for molecular LN staging reliably identifies CK19 mRNA in prostate
cancer. These results suggest that the OSNA assay may be suitable to improve (intraoperative) LN
staging in prostate cancer. For further verification, OSNA analysis of LN specimens from prostate cancer
patients is required.
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