HHE Pathology and diagnostics supplement 2018 - Page 12

generate false-positive signals, thus decreasing the overall accuracy. According to previous work, not all the detection targets are tumour specific, because the same targets can be found in blood cells. 45,46 Pantel et al have reported that CK-19, a major marker for CTC detection, is also present in immune cells. 47 Multiplex PCR, such as the AdnaTest kit (AdnaGen AG), could overcome this limitation. 45,48 Moreover, considering the genetic heterogeneity of CTCs, a multiplex PCR assay might provide improved sensitivity and specificity rates. Functional assays Functional assays exploit aspects of live cellular activity for CTC detection. Interestingly, these assays have the particularity to focus on the discovery of ‘metastasis competent cells’. In order to detect only viable CTCs, the functional Epithelial ImmunoSPOT (EPISPOT) assay, was introduced for in vitro CTC detection. 49 This technology assesses the presence of CTCs based on secretion, shed or release of specific proteins during 24–48 hours of short-term culture. 50 EPISPOT has been applied to blood and bone marrow samples, and validated in several different cancers, for example, breast and colon with the CK19 EPISPOT assay, or prostate with the PSA-EPISPOT assay. 51–54 This test is currently being further developed into a liquid format with micro-droplets (called EPIDROP) that allows capture of single viable CTCs and subsequent molecular characterisation. Another in vitro functional assay, Vita-Assay™ (Vitatex), exploits the preferential adhesion of invasive rare blood cells to a specialised matrix to enrich viable CTCs from blood up to one million- fold. 55 This method has also been tested in metastatic prostate 56 and ɕЁȸ)%٥ټхЁɵѥ)хɅхѥѥеɥٕ) Q ́ѼչЁ聵хх͕)ѡЁݕɔɽݸѕȁᕹɅхѥ)ɥ Q ́ٔѡЁɅѕɥѥ́)хх̵ͥѥѽȁ̸ɕЁѥ)ݥѠ͵չȁ͡ݕѡЁ Q )ɽѥ́ݥѠѡȁ͕ͥѥٔ)ɕɅѽյ́ɔյɥ)չɽ͕ѡɕձх) Q ɥٕ́ɽɕѡɕ͔)ѡȁѥЁѼѥմѽͥ)ѡɅ一!ݕٕȰЁɕ͕аѡ͔)٥ټ́ͅɕեɔٕ䁡 Q 奕́ѡ)Ʌѕͅݡٔͼ)䁉ٕ܁ѥ̸) Q ̰́ե)䰁)ѡѼ)չх)ѡ)ххͥ)ɽ٥)ɭȁ)ͽ͕)ɕѵЁ)ȁѥ)ɕѕЁɽѕѕɕи5ձѥ)́ͥЁՅɕɥѕѼ)ɽѕ́ѕɕЁȁյȁ䁅)ȁѡɅ丁Q́䁡Ѽѥ)ͥѡ݅́ɕمЁѼххͥ)ٕЁɕѵЁɕ̸͕%ɕ)ȁѥ̰ѡ!Hȁх́ Q ́ձ)͕͕͡܁͍ɕ́ݥѠɥ)յȁх̸5ɔɕѱ䰁չ)Ёɕձѽ́Ս́ɽɅ)Ѡāٔ፥ѥ)ѡɅѥхɝ́ձ͕ȁե)䁥ɔɥ́ѥ)չɝչЁİ)ȁ͕ͥՕѼѥ䁝)Ʌ͍ɥѽɅѕɥѥ́ Q ̰)Ց͕͕́ٔ́)ɥЁݡѥ̀]̤)Ѽɕ͔ѡչЁ9ݡ)Չ͕ՕѱՉѕѼѡ͕́)хѥ́䁹յȁمɥѥ́ͥ)ٕѥеɅѥ͕Օ)ѕ̸̰ܰЁ́ᅵ Q ́ݥѠ)хѕ-IĹݥ͍ѤHѡɅ)ѡȁɱ䁑ѕѥЁѼե)ѡɅ䁥٥Յѥ̰ѡ՝)ѕݽѡѡЁ]́ѡɕЁɥͬ)Ս́ѡɕձ́Ѽ)ѡɕɔɕձمѕ ͥ́ͽѥ)ͥ Q ̰ɥЁϊLЁչ)ЁՙЁѼхɅѥ) Q ̰ݡ́ѡ)ɅѡЁ́եхȁ͕ͥѥٔ)хѥ͕́ѕ́Ս́х)A H)ѡȁɽ́%M ͥ́ͥ) Q ́ѥ䁥չѽ一ذ)MՍչ%M ɽ)ݥѠѽѕѕѥ Q ́Ё)ͥȁѼЁɔѥ̸) ͥ)QɕЁͥѡ Q )́ɕѕѡɥ Q ѕ)ٕݥѡѡЁ̸9)ѕ́ٔɥ͕Ѽɕ́)́́ȁչх Q )ٽٕ́ɵѱ丁IЁ͕́ѡ)ѡѼ͕嵅Ʌͥѥѕ)ɭ́ Q ́յѕѡѕѥ)مՕ́܁ɭ́ Q ́ѡ䁵)ɽ٥ɵѥѕɕи)]Ց́ͥ Mɍ)ѡȁ Q ѕ́ٔɵѡЁ Q )յɅѥɽ٥́ɕمЁɽѥ)ɵѥم䰁ѡѕѥ)ȁեѼɕ́ѥ䁥́ѥ)չȁٕѥѥ%ͥե)ѥ́ЁѼ́ѡɕ)ȁ͍ɕѥ̰ݡձ)ѕѥɕՍѠɔ̸)MɅѕ́ȁ Q Ʌѕɥͅѥ) Q ́ѡѼչхѡ)ххͥ́ɽ٥ɭȁѼ)مٕͥ䁵ɔѡٽѥյ)ՉɥɕѵЁ͕͔)ɽɕͥ%ɽٕ́ѕ́Ѽ)奕ɕȁ Q ձѥ́Ё܁)ѕȁձȁձȁٕѥѥ̸) Ʌѕɥͅѥ Q ́́ѕȁͥ)Ѽյȁѕɽ䰁ݥѡЁ̰ͅ)Ցչսɕ͍Ʌ  )Ʌѥ͕ՕѠ9I9)սɕ͍ͥԁɥͅѥ%M )Aɽѕ͕́ͥ Q ́ɔɕѱ)əɵ䁥չхݥѠѥ)ݱ)Qѡ́ɕٕЁɽ=L)ѡ9ѥ%ѥє Ȁ%9 I )չѥ 9 H%%مѥٔ)5́%ѥѥٔ)ЁUхչ)ɅЁɕЁ䰁͕)ɥѥ́ɽѡɽUéMٕѠ)ɅݽɬAɽɅ@ܼܴ̤A%)ϊdɥѥ()!!хѡɔ