Digital PCR for Minimal Residual Disease Quantitation Using Immunoglobulin/T-Cell Receptor Gene Rearrangements in Acute Lymphoblastic Leukemia

Lü, Yi; Li, Zhenhua; Lim, Evelyn Huizi; Huan, Pei Tee; Kham, Shirley Kow Yin; Yeoh, Allen Eng Juh

doi:10.1016/j.jmoldx.2022.03.004

Cited by 1 publication

(2 citation statements)

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“…3 Lu et al propose an objective, statistics-based analytic algorithm to interpret raw data gained by using ddPCR. 17 Their study a well as our findings specifically relate to childhood ALL, but technical aspects for MRD determination as well as interpretation guidelines are of great importance for the MRD determination of many hematologic diseases.…”

Section: Non-specific Amplification Can Still Represent a Problem For...mentioning

confidence: 82%

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Transferring measurable residual disease measurement in pediatric acute lymphoblastic leukemia from quantitative real‐time PCR to digital droplet PCR

Luib

Kreyenberg

Michaelis

et al. 2023

Pediatric Transplantation

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Background Since the measurement of measurable residual disease (MRD) is part of clinical routine examination for children affected with acute lymphoblastic leukemia (ALL), continuous efforts are made to improve its method, applicability and accuracy. Whereas quantitative real‐time polymerase chain reaction (qPCR) is considered as the gold standard for MRD detection and endowed with international guidelines for implementation and evaluation, these do not yet exist for digital droplet PCR (ddPCR). However, advantages are seen in droplet partitioning for MRD measurement to allow absolute quantification without depending on reference samples. Methods In this study, 17 MRD targets of nine patients with childhood B‐ALL were analyzed with qPCR and ddPCR, respectively. All patients were assigned to high risk group and had hematopoietic stem cell transplantation and CD19 antibody therapy for relapse prevention. Starting with the sequences and guidelines of qPCR and optimizing the protocol for ddPCR, the MRD targets could also be measured precisely with this novel method, using the same primer and probe sets as for qPCR. Results The already established MRD protocol of qPCR could be transferred to ddPCR and all 17 MRD targets were measured in dilution series reaching comparable Limit of detection levels with both PCR methods. Conclusions With a given qPCR protocol and some experience in conventional MRD monitoring, it is conceivable to transfer the procedure of MRD measurement to ddPCR technology. Our data is in line with other studies which are summarized and discussed here as well to facilitate the transfer of MRD diagnostics to ddPCR.

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Section: Non-specific Amplification Can Still Represent a Problem For...mentioning

confidence: 82%

“…Even though different groups established ddPCR assays for various hematologic malignancies, they all concluded that ddPCR was well suited for MRD determination 3–27 . For a detailed overview about different ddPCR approaches for MRD detection, see Table 1.…”

Section: Discussionmentioning

confidence: 99%