Novel method for rapid fluorescence in-situ hybridization of ALK rearrangement using non-contact alternating current electric field mixing

Satoshi Fujishima,Kazuhiro Imai,Satoru Motoyama,Kaori Terata,Yoichi Akagami,Yoshitaro Saito,Hajime Saito,Ryuta Nakamura,Yusuke Sato,Hiroshi Nanjo,Yoshihiro Minamiya

doi:10.1038/s41598-017-15515-1

Satoshi Fujishima, Kazuhiro Imai + Show 9 more

Open Access

https://doi.org/10.1038/s41598-017-15515-1

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Abstract

Echinoderm microtubule-associated protein-like 4 gene and anaplastic lymphoma kinase gene (EML4-ALK) rearrangement is a key driver mutation in non-small cell lung cancer (NSCLC). Although Break-Apart ALK fluorescence in situ hybridization (FISH) is a reliable diagnostic method for detecting ALK gene rearrangement, it is too costly and time-consuming for use as a routine screening test. Our aim was to evaluate the clinical utility of a novel rapid FISH (RaFISH) method developed to facilitate hybridization. RaFISH takes advantage of the non-contact mixing effect of an alternating current (AC) electric field. Eighty-five specimens were used from patients diagnosed with NSCLC identified immunohistochemically as ALK 0, (1/2+) or (3+). With RaFISH, the ALK test was completed within 4.5 h, as compared to 20 h needed for the standard FISH. Although RaFISH produced results more promptly, the staining and accuracy of the ALK evaluation with RaFISH was equal to the standard. We found 97.6% agreement between FISH and RaFISH based on the status of the ALK signals. These results suggest RaFISH could be used as a clinical tool to promptly determine ALK status.

Highlights

Several key genetic alterations that drive non-small cell lung cancer (NSCLC) have been detected
We demonstrated in patients with breast cancer that rapid dual in-situ hybridization performed with an alternating current (AC) electric field and this device could be used to detect human epidermal growth factor receptor 2 (HER2) amplification within 6 h18
rapid FISH (RaFISH) enabled detection of ALK break apart within 4.5 h, which is much less time than the 20 h required for standard fluorescence in situ hybridization (FISH) (Table 2)

Summary

Introduction

Several key genetic alterations that drive non-small cell lung cancer (NSCLC) have been detected These include mutation of the epidermal growth factor receptor (EGFR) gene and rearrangement of the echinoderm microtubule-associated protein-like 4 gene and anaplastic lymphoma kinase gene (EML4-ALK)[1,2,3,4]. Because ALK gene rearrangement involves large chromosomal inversion and translocation, fluorescence in situ hybridization (FISH) has become the method of choice for detecting all forms of ALK gene rearrangement, including the rare non-EML4 fusions, and it was the assay used to detect this genetic aberration in the first clinical trials of the ALK inhibitor therapy with Crizotinib[8,10,11]. The aim of the present study was to evaluate the clinical utility, reliability and sensitivity of a novel rapid FISH (RaFISH) method that promotes ALK break-apart hybridization by taking advantage of the non-contact mixing effect in microdroplets subjected to an AC electric field

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