Effectiveness of digital PCR for MYD88L265P detection in vitreous fluid for primary central nervous system lymphoma diagnosis

Abstract

Primary central nervous system lymphoma (PCNSL) is a rare type of primary extranodal lymphoma (PEL). MYD88 L265P mutation has been observed in up to 75% of PCNSL cases, however, the validity and sensitivity of digital PCR in detecting this mutation remains to be elucidated. A total of 44 PCNSL patients, 15 diffuse large B-cell lymphoma not otherwise specified (DLBCL-NOS) patients and 13 other PEL patients were enrolled in the present study. The abilities of reverse transcription quantitative PCR (RT-qPCR) and droplet digital PCR (ddPCR) to detect the MYD88 L265P mutation in cerebral spinal fluid (CSF) samples were compared. The results suggested that ddPCR showed superior mutation detection sensitivity when compared with RT-qPCR (58 vs. 15%; P<0.05). The MYD88 L265P mutation was significantly associated with increased MYD88 protein overexpression in PCNSL brain tissue samples (P<0.05). Analysis of MYD88 L265P mutation status in CSF and vitreous fluid samples using ddPCR may be a promising technique for minimally invasive confirmation of PCNSL diagnosis.

EBV-Positive Gastric Cancer: Current Knowledge and Future Perspectives

“…Droplet digital PCR (ddPCR) is the latest method that can be used in the detection of Plasmodium falciparum ( 24 ), multiple viruses ( 25 ), nervous system lymphomas ( 26 ), etc. A ddPCR-based screening method for detection of EBV–associated gastric carcinoma was established in 2019 ( 27 , 28 ).…”

Digital Droplet PCR in Hematologic Malignancies: A New Useful Molecular Tool

“…In 2018, our group contributed to set ddPCR for the identification of the MYD88 mutation; this technique detected mutation in 96% of MW and 87% of IgM-MGUS cases (vs 81% and 58% by QT-PCR, respectively); the concordance rate with QT-PCR amounted to 78% on bone marrow and 68% on peripheral blood samples; in the remaining cases, ddPCR confirmed its advantage. The most interesting finding of this work was the possible application of this molecular tool to the circulating tumor DNA (ctDNA) harvested from plasma [ 88 ] or in the cerebral spinal fluid [ 89 ]. In 2019, another group employed ddPCR for detecting MYD88L265P mutation in a cohort of 39 patients; with a sensitivity of 1 × 10 −3 , the authors identified the mutation in 90% of MW cases, in 44% of patients affected by LPL, in 5% of IgM MM, and no in CLL or mantle cell lymphoma (MCL) cases [ 90 ].…”

cfDNA Sequencing: Technological Approaches and Bioinformatic Issues

“…As an example in lymphoma, this technique has a potential clinical use in diffuse large B cell lymphoma (DLBCL), as co-occurring mutations in MYD88 and CD79B can predict response to Ibrutinib treatment, thus providing a predictive molecular tool for patient and therapy selection [ 46 ]. As well, in primary central nervous system lymphoma (PCNSL), mutation MYD88 L265P was identified by ddPCR in cerebrospinal fluid or vitreous fluid with a superior sensitivity when compared with qPCR [ 47 , 48 ]. Since this mutation is found in up to 85% of PCNSL cases and not in non-hematological brain tumors, this ddPCR assay may be a promising technique for minimally invasive confirmation of PCNSL diagnosis.…”