Table 1.

Methodology for MRD detection

Method	Sensitivity	Quantitative (range)	Standardized	Sample requirements	Influences on assay performance9
FLC CD5/19 κ/λ2,6,22	10−2	No	No	Live leukocytes*	Number of benign B cells
Quantitative 4-color FLC7,8,27	10−4-10−5	Yes, to 10−4	Yes7	≥107 live leukocytes*	Number of available leukocytes; sensitivity reduced in paucicellular specimens
Consensus IGHV PCR27	∼10−3	No	Yes	∼106 total leukocytes†	Number of benign B cells, clone
ASO IGHV RQ-PCR33-35	10−5	Yes, to 10−4	Yes35	∼106 total leukocytes†	Clone (CDR3 region)
Nested ASO IGHV PCR36	10−6	No	No	∼106 total leukocytes†	Clone (CDR3 region)
Consensus IGHV PCR using HTS37,38	10−6	Yes, to 10−5	No	1 μg DNA for baseline sample, 6-60 μg for follow-up†	Specific sequences may amplify poorly in mutated CLL

^*Fresh, tested within 48 hours of collection or cryopreserved in liquid nitrogen. Live cells with intact membrane required.

^†DNA is stable at room temperature for prolonged periods of time, allowing these tests to be performed on historical samples. Live cells are not required.