title: [Clinical utility of real-time fluorescent PCR for combined detection of anaplastic lymphoma kinase and c-ros oncogene 1 receptor tyrosine kinase in non-small cell lung cancer].
abstract: Objective:To investigate the clinical application value of combined detection of ALK fusion gene and c-ros oncogene 1 receptor tyrosine kinase (ROS1) fusion gene in non-small cell lung cancer (NSCLC) using real-time fluorescent PCR.Methods:A kit for combined detection of ALK fusion gene and ROS1 fusion gene based on fluorescent PCR was used to simultaneously detect the two fusion genes in 302 cases of NSCLC specimens. The results were validated through Sanger sequencing. The consistency of the two detection methods was analyzed.Results:All 302 cases of NSCLC specimens were successfully analyzed through fluorescent PCR (302/302). 12 cases (4.0%) were found to contain ALK fusion gene, including 3 cases with ALK-M1, 3 with ALK-M2, 3 with ALK-M3, 1 with ALK-M4, and 2 with ALK-M6 fusion gene.12 cases (4.0%) were found to contain ROS1 fusion gene, including 1 case with ROS1-M7, 8 cases with ROS1-M8, 1 case with ROS1-M12, 1 case with ROS1-M14, and 1 case with double-positive ROS1-M3 and ROS1-M8 fusion genes. The total detection rate of ALK fusion gene and ROS1 fusion gene was 7.9% (24/302) and 278 cases showed to be negative for ALK fusion gene and ROS1 fusion gene. The successful detection rates for Sanger DNA sequencing were also 100%. The positive, negative and total coincidence rates obtained by real-time fluorescent PCR and by Sanger DNA sequencing were all 100%.Conclusions:The results of Sanger DNA sequencing demonstrate that the real-time fluorescent PCR assay is equally effective in detecting ALK and ROS1 fusion genes in NSCLC tissues. Furthermore, real-time fluorescent PCR assay can be used to detect trace ALK and ROS1 fusion gene simultaneously in tiny samples, and can save time and avoid repeated sampling. It is worthy of recommendation as a rapid and reliable detection technique.
authors.affiliation: Department of Pathology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: D Y
authors.initials: DY
authors.lastname: Bai
authors.affiliation: Department of Pathology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: H P
authors.initials: HP
authors.lastname: Zhang
authors.affiliation: Department of Pathology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: S
authors.initials: S
authors.lastname: Zhong
authors.affiliation: Department of Pathology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: W H
authors.initials: WH
authors.lastname: Suo
authors.affiliation: Department of Pathology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: D H
authors.initials: DH
authors.lastname: Gao
authors.affiliation: Department of Pathology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Department of Pathology, the First Affiliated Hospital of Xiamen University, Xiamen 361003, China.
authors.forename: J H
authors.initials: JH
authors.lastname: Tu
mesh_terms: D000077548:Anaplastic Lymphoma Kinase; D002289:Carcinoma, Non-Small-Cell Lung / Q000201:enzymology*; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology*; D015514:Oncogene Proteins, Fusion / Q000032:analysis*; D009857:Oncogenes; D011505:Protein-Tyrosine Kinases / Q000032:analysis*; D011518:Proto-Oncogene Proteins / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020794:Receptor Protein-Tyrosine Kinases / Q000032:analysis* / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D015514:Oncogene Proteins, Fusion; D011518:Proto-Oncogene Proteins; C000626173:ALK protein, human; D000077548:Anaplastic Lymphoma Kinase; D011505:Protein-Tyrosine Kinases; C062333:ROS1 protein, human; D020794:Receptor Protein-Tyrosine Kinases