同源重组修复检测的评价

Evaluation of homologous recombination repair detection

目的使用人实体瘤多基因突变联合检测试剂盒,对国家参考品进行同源重组修复基因(HRR)检测,评价试剂盒的HRR检测能力。方法使用超声打断仪将参考品基因组DNA打断到180~220 bp。将片段化DNA进行末端修复、加接头和PCR扩增等步骤后制备预文库;然后杂交捕获后获得终文库;最后使用NextSeq550Dx测序仪进行测序。结果国家参考品标示的BRCA基因突变位点以及突变的外显子位置均能准确检出,而且没有检出其他非标示的致病性或疑似致病性突变位点。试剂盒虽然对部分BRCA基因突变位点的解读结果与标示的解读结果有差异,但是解读结果也符合国家参考品的准确性要求。试剂盒未对其他的HRR基因突变进行解读。结论HRR检测对BRCA基因突变按照BRCA基因解读规则进行解读,对其他的HRR基因突变的解读结果需要制定相应的解读规则。

Objective To detect homologous recombinant repair genes(HRR)in national material reference using the human solid tumor multi-gene mutation detection kit and evaluate the detection capability.Methods Genomic DNA was interrupted to about 180-220 bp by ultrasonic interrupter.The fragment DNA was prepared by terminal repair,splicing and PCR amplification.Then the library was obtained after hybridization and capture.Finally,NextSeq550 Dx sequencer was used for sequencing.Results The mutation sites and exon positions of BRCA gene marked by national reference materials can be accurately detected,and no other non marked pathogenic or suspected pathogenic mutation sites have been detected.Although the interpretation results of some BRCA gene mutation sites by the kit are different from the labeled interpretation results,but the interpretation results also meet the accuracy requirements of national reference materials.The kit does not interpret other HRR gene mutations.Conclusion BRCA gene mutations should be interpreted according to BRCA gene interpretation rules,while the interpretation of other HRR gene mutations requires the formulation of corresponding interpretation rules.