Combination of conventional immunohistochemistry and qRT-PCR to detect ALK rearrangement

Abstract

Background: Compared with FISH and qRT-PCR analyses, immunohistochemistry (IHC) is the preferred screening test in most pathology practices for ALK-rearrangement detection. With 100% sensitivity and 98% specificity, the VENTANA ALK (D5F3) IHC assay has been approved in the EU and some Asian countries for ALK-rearrangement detection. However, an automated Ventana IHC platform is not available in most pathology labs. In this study, we evaluated the applicability of conventional IHC with D5F3 antibody in routine pathological practice and proposed detection methods and procedures that ensure that patients with ALK+ are not missed. Methods: FISH and IHC analyses were performed on 297 lung adenocarcinoma cases. VENTANA IHC and qRT-PCR assay were applied to evaluate ALK-fusion status in the discordant cases of FISH and IHC. The association of ALK+ with clinicopathological characteristics was statistically analyzed.

Clinical characteristics of patients with non-small cell lung cancers harboring anaplastic lymphoma kinase rearrangements and primary lung adenocarcinoma harboring epidermal growth factor receptor mutations

“…The incidence of ALK rearrangements was found to be 9.6% (73/763) in NSCLC and 11.0% (54/493) in adenocarcinoma in our study, which was similar to the results previously published in Chinese (Shan et al, 2014;Wang et al, 2014a;Yang et al, 2014) and Asian cohorts (Kim et al, 2014). As previously reported, patients who were ALK-positive were predominantly young women and never/light smokers (Lynch et al, 2004;Rodig et al, 2009;Shaw et al, 2009;Wong et al, 2009;Jokoji et al, 2010;Takahashi et al, 2010;Zhang et al, 2013;Zhang et al, 2014).…”

“…Recently many studies have reported the efficacy of Ventana IHC for detecting ALK rearrangements with D5F3 antibodies and have supported that Ventana IHC is a method with excellent sensitivity and specificity to identify ALK rearrangements in paraffin-embedded lung cancer specimens (Takahashi et al, 2010;Zhang et al, 2013;Shan et al, 2014;Wang et al, 2014a;Zhang et al, 2014). The sensitivity and specificity of Ventana IHC were found to be 100 and 98.2%, respectively, and the concordance rate between the fluorescence in situ hybridization (FISH) and the Ventana IHC was 98.4% (Wang et al, 2014a).…”

“…The sensitivity and specificity of Ventana IHC were found to be 100 and 98.2%, respectively, and the concordance rate between the fluorescence in situ hybridization (FISH) and the Ventana IHC was 98.4% (Wang et al, 2014a). The 100% sensitivity measure suggested that IHC analysis is an effective way to screen patients in the clinical diagnosis process (Shan et al, 2014). Although break-apart FISH is the standard method for diagnosis of ALK rearrangements, it is expensive and not readily available.…”

“…Immunohistochemistry (IHC) for ALK protein overexpression is a promising screening modality, with D5F3 antibodies showing excellent sensitivity and specificity for ALK-rearranged NSCLC (Takahashi et al, 2010;To et al, 2013;Shan et al, 2014;Zhang et al, 2013;. Furthermore, the ALK fusion protein IHC diagnostic kit developed by Roche Ventana Medical Systems (Oro Valley, AZ, USA) has been approved by the European Medicines Agency (EMEA) and the China Food and Drug Administration (CFDA) as an aid for identifying patients who are eligible for treatment with crizotinib.…”

Diagnosis of EML4 - ALK Translocation With FISH, Immunohistochemistry, and Real-time Polymerase Chain Reaction in Patients With Non–Small Cell Lung Cancer

“…This assay has been approved for ALK-rearrangement detection by the FDA in China and Taiwan. [23][24][25] From the…”