Int. J. Mol. Sci. 2013, 14(7), 12931-12952; doi:10.3390/ijms140712931
Article

Metasin—An Intra-Operative RT-qPCR Assay to Detect Metastatic Breast Cancer in Sentinel Lymph Nodes

1 Cellular Pathology, Princess Alexandra Hospital NHS Trust, Harlow, Essex CM20 1QX, UK 2 Breast Unit, Prince Philip Hospital Bryngwynmawr, Llanelli Carmarthenshire SA14 8QF, UK 3 Department of Histopathology, Southend University Hospital NHS Trust, Southend, Essex SS0 0RY, UK 4 Department of Pathology, Jules Bordet Institute, Brussels 1000, Belgium 5 Postgraduate Medical Institute, Faculty of Health, Social Care and Education, Anglia Ruskin University, Chelmsford, Essex CM1 1SQ, UK
* Author to whom correspondence should be addressed.
Received: 27 March 2013; in revised form: 1 May 2013 / Accepted: 15 May 2013 / Published: 24 June 2013
(This article belongs to the Section Molecular Diagnostics)
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Abstract: Nodal status is one of the most important prognostic factors in breast cancer. Established tests such as touch imprint cytology and frozen sections currently used in the intra-operative setting show variations in sensitivity and specificity. This limitation has led to the development of molecular alternatives, such as GeneSearch, a commercial intra-operative real-time quantitative Polymerase Chain Reaction (RT-qPCR) assay that allows the surgeon to carry out axillary clearance as a one-step process. Since GeneSearch has been discontinued, we have developed the replacement Metasin assay, which targets the breast epithelial cell markers CK19 and mammaglobin mRNA and identifies metastatic disease in sentinel lymph nodes. The optimised assay can be completed within 32 min (6 min for RNA preparation and 26 min instrument run time), making its use feasible in the intraoperative setting. An analysis by Metasin of 154 archived lymph node homogenates previously analysed by both parallel histology and GeneSearch showed concordance for 148 cases. The sensitivity and specificity of Metasin compared with GeneSearch were 95% (CI 83%–99%) and 97% (CI 91%–99%) respectively; compared with histology they were 95% (CI 83%–99%) and 97% (CI 91%–99%), respectively. The sensitivity and specificity of GeneSearch compared with histology were 90% (CI 77%–96%) and 97% (CI 93%–99%) respectively. The positive predictive value of Metasin was 90% and negative predictive value was 98% for both histology and GeneSearch. The positive predictive value of GeneSearch was 92% and the negative predictive value was 97% compared to histology. The discordance rates of Metasin with both GeneSearch and histology were 3.89%. In comparison, the discordance rate of GeneSearch with histology was 4.5%. Metasin’s robustness was independently evaluated on 193 samples previously analysed by GeneSearch from the Jules Bordet Institute, where Metasin yielded comparable results.
Keywords: Metasin; CK19; sentinel node; metastatic; GeneSearch; intra-operative; OSNA; NICE; Mammaglobin; Cepheid; qPCR; POC; breast; cancer; PBGD; axillary; immunostains; MGB

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MDPI and ACS Style

Al-Ramadhani, S.; Sai-Giridhar, P.; George, D.; Gopinath, P.; Arkoumani, E.; Jader, S.; Sundaresan, M.; Salgado, R.; Larsimont, D.; Bustin, S.A.; Sundaresan, V. Metasin—An Intra-Operative RT-qPCR Assay to Detect Metastatic Breast Cancer in Sentinel Lymph Nodes. Int. J. Mol. Sci. 2013, 14, 12931-12952.

AMA Style

Al-Ramadhani S, Sai-Giridhar P, George D, Gopinath P, Arkoumani E, Jader S, Sundaresan M, Salgado R, Larsimont D, Bustin SA, Sundaresan V. Metasin—An Intra-Operative RT-qPCR Assay to Detect Metastatic Breast Cancer in Sentinel Lymph Nodes. International Journal of Molecular Sciences. 2013; 14(7):12931-12952.

Chicago/Turabian Style

Al-Ramadhani, Salma; Sai-Giridhar, Priya; George, Dilushana; Gopinath, Preethi; Arkoumani, Evdokia; Jader, Samar; Sundaresan, Maryse; Salgado, Roberto; Larsimont, Dennis; Bustin, Stephen A.; Sundaresan, Vasi. 2013. "Metasin—An Intra-Operative RT-qPCR Assay to Detect Metastatic Breast Cancer in Sentinel Lymph Nodes." Int. J. Mol. Sci. 14, no. 7: 12931-12952.

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