Accurate detection of altered anaplastic lymphoma kinase (ALK) expression is critical for the selection of lung cancer patients eligible for ALK-targeted therapies. To overcome intrinsic limitations and discrepancies of currently available companion diagnostics for ALK, we developed a simple, affordable and objective PCR-based predictive model for the quantitative measurement of any ALK fusion as well as wildtype ALK upregulation. This method, optimized for low-quantity/-quality RNA from FFPE samples, combines cDNA pre-amplification with ad hoc generated calibration curves. All the models we derived yielded concordant predictions when applied to a cohort of 51 lung tumors, and correctly identified all 17 ALK FISH-positive and 33 of the 34 ALK FISH-negative samples. The one discrepant case was confirmed as positive by IHC, thus raising the accuracy of our test to 100%. Importantly, our method was accurate when using low amounts of input RNA (10 ng), also in FFPE samples with limited tumor cellularity (5-10%) and in FFPE cytology specimens. Thus, our test is an easily implementable diagnostic tool for the rapid, efficacious and cost-effective screening of ALK status in patients with lung cancer.

Sensitive and affordable diagnostic assay for the quantitative detection of anaplastic lymphoma kinase (ALK) alterations in patients with non-small cell lung cancer / Dama, E; Tillhon, M; Bertalot, G; de Santis, F; Troglio, F; Pessina, S; Passaro, A; Pece, S; de Marinis, F; Dell'Orto, P; Viale, G; Spaggiari, L; Di Fiore, Pp; Bianchi, F; Barberis, M; Vecchi, M. - In: ONCOTARGET. - ISSN 1949-2553. - 7:24(2016), pp. 37160-37176. [10.18632/oncotarget.9471]

Sensitive and affordable diagnostic assay for the quantitative detection of anaplastic lymphoma kinase (ALK) alterations in patients with non-small cell lung cancer

Bertalot G;
2016-01-01

Abstract

Accurate detection of altered anaplastic lymphoma kinase (ALK) expression is critical for the selection of lung cancer patients eligible for ALK-targeted therapies. To overcome intrinsic limitations and discrepancies of currently available companion diagnostics for ALK, we developed a simple, affordable and objective PCR-based predictive model for the quantitative measurement of any ALK fusion as well as wildtype ALK upregulation. This method, optimized for low-quantity/-quality RNA from FFPE samples, combines cDNA pre-amplification with ad hoc generated calibration curves. All the models we derived yielded concordant predictions when applied to a cohort of 51 lung tumors, and correctly identified all 17 ALK FISH-positive and 33 of the 34 ALK FISH-negative samples. The one discrepant case was confirmed as positive by IHC, thus raising the accuracy of our test to 100%. Importantly, our method was accurate when using low amounts of input RNA (10 ng), also in FFPE samples with limited tumor cellularity (5-10%) and in FFPE cytology specimens. Thus, our test is an easily implementable diagnostic tool for the rapid, efficacious and cost-effective screening of ALK status in patients with lung cancer.
2016
24
Dama, E; Tillhon, M; Bertalot, G; de Santis, F; Troglio, F; Pessina, S; Passaro, A; Pece, S; de Marinis, F; Dell'Orto, P; Viale, G; Spaggiari, L; Di F...espandi
Sensitive and affordable diagnostic assay for the quantitative detection of anaplastic lymphoma kinase (ALK) alterations in patients with non-small cell lung cancer / Dama, E; Tillhon, M; Bertalot, G; de Santis, F; Troglio, F; Pessina, S; Passaro, A; Pece, S; de Marinis, F; Dell'Orto, P; Viale, G; Spaggiari, L; Di Fiore, Pp; Bianchi, F; Barberis, M; Vecchi, M. - In: ONCOTARGET. - ISSN 1949-2553. - 7:24(2016), pp. 37160-37176. [10.18632/oncotarget.9471]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11572/360238
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