ROS1融合基因检测方法的建立

Establishment of ROS1 gene fusion testing method

目的:建立检测ROS1融合基因的荧光PCR法,为肺癌病人的个体化药物治疗提供新的检测试剂盒。方法:根据人类ROS1基因的外显子和融合伙伴基因的外显子序列设计引物和探针,建立ROS1融合基因的荧光PCR法。然后采用不同ROS1基因融合的假病毒质粒作为质控品,评价方法的准确性、检测限和特异性。检测350例临床样本,并与直接测序结果进行比较,评价方法的临床适用性。结果:建立的ROS1融合基因PCR-荧光探针法,能准确检测出14种不同的ROS1融合基因;检测限是不高于总量0.01μg的RNA模板;正常人RNA的结果显示为融合突变阴性。临床样本的结果与测序结果的一致性好,Kappa值为1.00。结论:建立的ROS1融合基因检测方法具有很好的准确性、检测限和特异性,并且具有很好的临床适用性。

Objective：To establish a fluorescence PCR method for ROS1 gene fusion detection,so as to develop a new detection kit for personalized targeted therapy for lung cancer.Methods：According to exon sequences of ROS1 and fusion partners,the primers and probes were designed.The fluorescence PCR method for ROS1 gene fusion detection was established.The pseudotyped virus plasmids with different ROS1 gene fusion were chosen as control materials for evaluating the accuracy,limit and specificity of the method.350 cases of clinical samples were detected to evaluate the clinical applicability.The results were compared with the direct sequencing results of these samples.Results：Fluorescence PCR method for ROS1 gene fusion detection could accurately detect14 kinds of ROS1 gene fusion.The limit was 0.01 μg total RNA template.The result of normal sample was not ROS1 gene fusion.The results of clinical samples showed the good consistency with the sequencing results,and the Kappa value was 1.00.Conclusion：Fluorescence PCR method for ROS1 gene fusion detection was proved to be an effective method with better accuracy,limit,specificity and clinical applicability.