{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,10]],"date-time":"2025-12-10T08:56:45Z","timestamp":1765357005708,"version":"3.37.3"},"reference-count":33,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2021,9,24]],"date-time":"2021-09-24T00:00:00Z","timestamp":1632441600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004663","name":"ministry of science and technology, taiwan","doi-asserted-by":"publisher","award":["110-2320-B-006-026"],"award-info":[{"award-number":["110-2320-B-006-026"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004844","name":"national cheng kung university hospital","doi-asserted-by":"publisher","award":["NCKUH-11008004"],"award-info":[{"award-number":["NCKUH-11008004"]}],"id":[{"id":"10.13039\/501100004844","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004844","name":"national cheng kung university hospital","doi-asserted-by":"publisher","award":["NCKUH-11008014"],"award-info":[{"award-number":["NCKUH-11008014"]}],"id":[{"id":"10.13039\/501100004844","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004844","name":"national cheng kung university hospital","doi-asserted-by":"publisher","award":["NCKUH-11008018"],"award-info":[{"award-number":["NCKUH-11008018"]}],"id":[{"id":"10.13039\/501100004844","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Approximately 5\u20137% of non\u2013small cell lung cancer (NSCLC) cases harbor an anaplastic lymphoma kinase (<jats:italic>ALK<\/jats:italic>) fusion gene and may benefit from <jats:italic>ALK<\/jats:italic> inhibitor therapy. To detect <jats:italic>ALK<\/jats:italic> fusion genes, we developed a novel test using reverse transcription polymerase chain reaction (RT-PCR) for the <jats:italic>ALK<\/jats:italic> kinase domain (KD). Since <jats:italic>ALK<\/jats:italic> expression is mostly silenced in the adult with the exception of neuronal tissue, the normal lung tissue, mesothelial lining, and inflammatory cells are devoid of <jats:italic>ALK<\/jats:italic> transcript, making <jats:italic>ALK<\/jats:italic> KD RT-PCR an ideal surrogate test for <jats:italic>ALK<\/jats:italic> fusion transcripts in lung or pleural effusion. The test was designed with a short PCR product (197 bp) to work for both malignant pleural effusion (MPE) and formalin-fixed, paraffin-embedded (FFPE) NSCLC samples. Using ALK IHC as a reference, the sensitivity of the test was 100% for both MPE and FFPE. The specificity was 97.6% for MPE and 97.4% for FFPE. Two false positive cases were found. One was a metastatic brain lesion which should be avoided in the future due to intrinsic <jats:italic>ALK<\/jats:italic> expression in the neuronal tissue. The other one resulted from <jats:italic>ALK<\/jats:italic> gene amplification. Due to potential false positivity, subsequent confirmation tests such as fluorescence in situ hybridization or multiplex PCR would be preferable. Nevertheless, the test is simple and inexpensive with no false negativity, making it a desirable screening test. It also offers an advantage over multiplex RT-PCR with the capability to detect novel <jats:italic>ALK<\/jats:italic> fusions. Indeed through the screening test, we found a novel <jats:italic>ALK<\/jats:italic> fusion partner (sperm antigen with calponin homology and coiled-coil domains 1 like gene, <jats:italic>SPECC1L<\/jats:italic>) with increased sensitivity to crizotinib <jats:italic>in vitro<\/jats:italic>. In summary, a novel RNA-based <jats:italic>ALK<\/jats:italic> KD analysis was developed for <jats:italic>ALK<\/jats:italic> rearrangement screening in MPE and FFPE specimens of NSCLC. This simple inexpensive test can be implemented as routine diagnostics.<\/jats:p>","DOI":"10.1371\/journal.pone.0257152","type":"journal-article","created":{"date-parts":[[2021,9,24]],"date-time":"2021-09-24T17:52:34Z","timestamp":1632505954000},"page":"e0257152","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Development of a novel ALK rearrangement screening test for non\u2013small cell lung 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