Accurate results
for patient care
Laboratory medicine and in vitro diagnostics
JCTLM Database: higher-order reference materials, methods and services
Details : C18RMP4 - NIM Reference measurement procedure for ''EGFR'' 19DEL mutation fraction abundance
| Type (acronym) | rmp |
|---|---|
| Type | reference measurement procedure |
| Unique Nomination Number | C18RMP4 |
| Analyte Name | Single nucleotide variant (EGFR 19DEL) |
| Method capable of Traceability of Analyte to SI or Defined Procedure | SI-traceable |
| Name | NIM Reference measurement procedure for EGFR 19DEL mutation fraction abundance |
| Organization that developed Validated Method | National institute of metrology, China |
| Measurement Techniques Used | Digital PCR |
| Comments 1 | The reference measurement procedure gives EGFR 19DEL muntant fractional abundance of the total EGFR gene. |
| Comments 2 | We have participated an international comparison, CCQM P184 which is about the BRAF V600E and EGFR 19DEL measurement and satisfied result was obtained by using our proposed RMP |
| Quantity | Amount-of-substance ratio (DNA copy number ratio) |
| From (0) | 0.2 |
| To (0) | 100 |
| Unit (0) | % |
| From (1) | 25 |
| To (1) | 3.2 |
| Unit (1) | % |
| Level of Confidence (%) | 95% |
| Comments on Concentration Range | The reference measurement procedure gives EGFR 19DEL mutant fractional abundance. The unit is % of EGFR 19DEL mutant fractional abundance in total EGFR gene. |
| Peer Reviewed Publication | Establishment of primary reference measurement procedures and reference materials for EGFR variant detection in non-small cell lung cancer, Wang X. et al, |
| Comments to be published via JCTLM DB | This reference measurement procedure (RMP) specifies the operating specifications for measurement of fractional abundance (FA) of DNA containing EGFR single nucleotide variant (SNV) or small deletion (DEL) mutation quantification by digital PCR, and is specifically designed for this SNV/DEL and their corresponding wild-type sequences. The RMP is only suitable for DNA samples which have been confirmed by sequencing to containing the SNV and DEL described HERE; it is not suitable for samples where the EGFR sequence variant is unknown. This method is applicable to the measurement of the three target EGFR FA in plasmid and genomic DNA in buffered solution. The RMP has been tested with materials containing fragmented gDNA[1],[2], however caution should be exercised when applying the RMP to fragmented DNA (<1 kb). The mutant and wild-type sequence-containing templates should have approximately the same fragment size distribution to avoid bias in the measurement of FA. DNA extracts from biological matrices should be checked for the presence of reaction inhibitors by checking purity by UV spectrophotometry and/or spiking methods[3]. It is recommended that plasmid DNA is linearised and high MW gDNA is subject to restriction digestion prior to dPCR[2]. The measurement interval of the RMP for FA is between 0.20% and 100%. The Limit of Detection (LoD) of L858R, 19DEL, and T790M mutation fractional abundance were determined to be 0.02%, 0.02%, and 0.06%, respectively, and the Limit of quantification (LoQ) for FA were determined to be 0.21%, 0.24% and 0.16%, respectively. [1] Establishment of Primary Reference Measurement Procedures and Reference Materials for EGFR variant detection in non-small cell lung cancer, Wang et al, Analytical Methods 2021 4 19 [2] Evaluation of droplet digital PCR and next generation sequencing for characterizing DNA reference material for KRAS mutation detection, Dong et al, Scientific Reports 2018 8 9650 [3] ISO 20395: 2019—Biotechnology—Requirements for evaluating the performance of quantification methods for nucleic acid target sequences— qPCR and dPCR. |
| Analyte Category Name | Nucleic acids |
| CCQM Key Comparison Report | CCQM P184 Standardisation of cell-free DNA measurements: An international study on comparability of low concentration DNA measurements using cancer variants
|
| Non-CCQM Key Comparison Report | See Anal. Methods 2021 13 2114-2123 for national interlaboratory study |
| Measurement principle/technique | Digital PCR |