The Application Of Unique Molecular Identifier Labeled Super Depth Next Generation Sequencing In Minimal Residential Disease Detection Of Pediatric Acute Leukemia

Objective: To establish the methods of minimal residential disease(MRD)detection in pediatric acute leukemia(AL)by using unique molecular identifier labeled next generation sequencing(NGS)technique and to analyse its sensitivity and specificity.Methods: Bone marrow samples were extracted in 35 cases of pediatric AL before and after remission induction.NGS technique was used for WES sequencing at diagnosis and biomarkers for MRD detection were chosen in patient specific tumor gene mutations.Such gene mutations were detected by unique molecular identifier(UMI)labeled super depth NGS technique and frequencies of mutated genes were recorded as MRD level.MRD markers were also seek by flow cytometry(FCM)and reversed transcriptase polymerase chain reactions(RT-PCR),MRD levels were also detected by FCM and PCR technique after remission induction,sensitivity and specificity of three methods were compared.Results: Positive rates of MRD marker detected by NGS technique were exceed than that by FCM and PCR technique(P<0.05);sensitivities of MRD monitoring by NGS technique were also Superior to FCM and PCR technique(P<0.05).There was no statistical difference for mean value of MRD levels which were measured by FCM and NGS technique(P>0.05).Conclusion: UMI labeled super depth NGS technique can be used for MRD monitoring in pediatric AL.