Cytology specimens offer an effective alternative to formalin-fixed tissue as demonstrated by novel automated detection for ALK break-apart FISH testing and immunohistochemistry in lung adenocarcinoma.

Minimally invasive sampling by cytology or core needle biopsy often provides an initial diagnosis for treatment in patients with lung nodules. From these limited specimens, multiple molecular studies are frequently requested. Current guidelines from the US Food and Drug Administration recommend using formalin-fixed paraffin-embedded tissue sections for the detection of anaplastic lymphoma kinase (ALK) gene rearrangement by fluorescence in situ hybridization (FISH). The authors compared alcohol-fixed and formalin-fixed cytology specimens using a novel automated detection for ALK rearrangements by FISH and immunohistochemistry (IHC). ALK FISH testing was performed on 129 lung adenocarcinomas from 71 cytology cases and 58 biopsy/resection specimens using Papanicolaou staining with integrated cytomorphology. IHC with the ALK D5F3 antibody was performed on cases with residual material (88 of 129 cases). The mean age of the patients was 66 years; there were 62 women and 67 men. ALK gene rearrangement was present in 4% of cytology specimens (3 of 71 specimens) and 7% of surgical specimens (4 of 58 specimens). FISH in 13 cases was technically unsuccessful. Of the 7 FISH-positive cases, only 2 cytology cases (4%) and 2 surgical cases (6%) were found to be positive with the ALK antibody, demonstrating 80% concordance. The one case found to be negative for ALK by IHC demonstrated a variant rearrangement of the ALK 2p23 gene locus by FISH. The results of the current study validate the usefulness of alcohol-fixed and/or formalin-fixed cytology specimens for ALK rearrangement by a novel automated FISH method. IHC using the D5F3 antibody for ALK is specific in this limited cohort. The authors also demonstrated that alcohol-fixed cytology specimens can be used for ALK rearrangement by automated FISH, alone or in conjunction with IHC.

Rosenblum F ，Hutchinson LM ，Garver J ，Woda B ，Cosar E ，Kurian EM ... -  《-》

ALK status testing in non-small-cell lung carcinoma by FISH on ThinPrep slides with cytology material.

Minca EC ，Lanigan CP ，Reynolds JP ，Wang Z ，Ma PC ，Cicenia J ，Almeida FA ，Pennell NA ，Tubbs RR ... -  《-》

Correlation of IHC and FISH for ALK gene rearrangement in non-small cell lung carcinoma: IHC score algorithm for FISH.

Yi ES ，Boland JM ，Maleszewski JJ ，Roden AC ，Oliveira AM ，Aubry MC ，Erickson-Johnson MR ，Caron BL ，Li Y ，Tang H ，Stoddard S ，Wampfler J ，Kulig K ，Yang P ... -  《-》

Screening of anaplastic lymphoma kinase rearrangement by immunohistochemistry in non-small cell lung cancer: correlation with fluorescence in situ hybridization.

Paik JH ，Choe G ，Kim H ，Choe JY ，Lee HJ ，Lee CT ，Lee JS ，Jheon S ，Chung JH ... -  《-》

Antibody 1A4 with routine immunohistochemistry demonstrates high sensitivity for ALK rearrangement screening of Chinese lung adenocarcinoma patients: A single-center large-scale study.

The rearrangement of echinoderm microtubule-associated protein-like 4-analplastic lymphoma kinase (EML4-ALK) in non-small cell lung cancer (NSCLC) cells might be a promising therapeutic target. However, the low positive rate seeks a reliable and cost-effective method for ALK rearrangement prescreening. This study aimed to evaluate the application of a novel primary antibody 1A4 for routine ALK immunohistochemistry (IHC) test. Primary antibody 1A4 and D5F3 were used for the screening of 595 formalin-fixed, paraffin-embedded tissues of consecutive patients with lung adenocarcinoma for ALK-positive candidates. Ventana detection system and fluorescence in-situ hybridization (FISH) were used as reference methods. Among 595 cases, the protein expression statuses of 1A4 were 3+ (18), 2+ (50), 1+ (153), and 0+ (374), and those of D5F3 were 3+ (17), 2+ (18), 1+ (20), and 0+ (540). Ventana detection system and FISH test results were successfully obtained from 482 cases. A total of 298 specimens with 1A4 (-) showed 100% concordance with standard FISH results. All 58 FISH (+) cases were identified by antibody 1A4. Meanwhile, 14 and 5 were missed by antibody D5F3 with routine IHC and Ventana system, respectively. 1A4 with routine IHC had better sensitivity (100%, 75.9%, and 91.4%, respectively), but lower specificity (70.3%, 99.8%, and 100%, respectively), than D5F3 with routine IHC and Ventana system. The novel antibody 1A4 used as a prescreening method may help to reduce the false-negative rearranged ALK status if FISH or reverse transcription polymerase chain reaction results were used for validation.

Wang Q ，Zhao L ，Yang X ，Wei S ，Zeng Y ，Mao C ，Lin L ，Fu P ，Lyu L ，Li Z ，Xiao H ... -  《-》