PCR (Polymerase Chain Reaction) primers, kit and liquid-phase chip for detecting ROS1 fused gene

Abstract

The invention discloses PCR (Polymerase Chain Reaction) primers, a kit and a liquid-phase chip for detecting an ROS1 fused gene. The liquid-phase chip comprises PCR amplification primers, an ASPE primer and a micro-ball, wherein the ASPE primer is composed of a tag sequence and specific primers; the sequences of the specific primers are as follows: a probe pair C6; SEQ ID NO.34 of R32 and the probe pair C6; SEQ ID NO.35 of R34 and a probe pair C5; SEQ ID NO.36 of the R34 and a probe pair S2; SEQ ID NO.37 of the R32 and a probe pair S4; SEQ ID NO.38 of the R32 and a probe pair S4; SEQ ID NO.39 of the R34 and a probe pair SL13; SEQ ID NO.40 of the R32 and a probe pair SL4; SEQ ID NO.41 of the R34 and a probe pair E10; SEQ ID NO.42 of the R34 and a probe pair L16; SEQ ID NO.43 of R35 and / or a probe pair T5; SEQ ID NO.44 of the R35. The liquid-phase chip has a very good signal-noise ratio and can finish the amplification of 11 fused subtypes in one step; the specific primer has very good specificity.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a PCR primer, a kit and a liquid phase chip for detecting ROS1 fusion gene. technical background [0002] v-ros UR2 sarcoma viral oncogene homolog 1 (v-ros avian UR2 sarcoma viral oncogene homolog 1, ROS1), in 1987 Rabin M. cloned the gene related to the growth of v-ros sarcoma and named it ROS gene. There are breakpoints at the 32nd, 34th and 35th exons of the ROS1 gene, which can be fused with various genes. The ROS1 fusion gene was confirmed as a new lung cancer driver gene, which plays an important role in the formation of lung adenocarcinoma. At the 48th annual meeting of the American Society of Clinical Oncology (ASCO) held in June 2012, Shaw of the Massachusetts General Hospital reported for the first time the phase I clinical trial of patients with ROS1 gene fusion, a new molecular target for lung cancer. The clinical trial proved t...

AI Technical Summary

Problems solved by technology

The RT-PCR method is to design primers for the known RET fusion gene types and fusion methods. After the RNA is reverse-transcribed to obtain cDNA, the target fragment is amplified by PCR for detection, which has low sensitivity and easy contamination of the sample. , the disadvantage of high false positive rate

Although the non-radioactive in situ hybridization technique FISH method is relatively intuitive, the test process is too cumbersome, requires a wide variety of reagents, is time-consuming and laborious, and it can only detect whether the fusion gene exists, and cannot accurately determine the specific fusion type. Sensitivity low, thus limiting the application of the law to a certain extent

Images