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Details : C18RMP12 - NIM Reference measurement procedure for SARS-CoV-2 ''N'' gene measurement
| Type (acronym) | rmp |
|---|---|
| Type | reference measurement procedure |
| Unique Nomination Number | C18RMP12 |
| Analyte Name | SARS-CoV-2 RNA N gene |
| Method capable of Traceability of Analyte to SI or Defined Procedure | SI traceable |
| Name | NIM Reference measurement procedure for SARS-CoV-2 N gene measurement |
| Organization that developed Validated Method | National institute of metrology, China |
| Measurement Techniques Used | Digital PCR |
| Comments 1 | The reference measurement procedure gives SARS-CoV-2 RNA copy number concentration |
| Comments 2 | We have participated an international comparison, CCQM P199B which is about the SARS-CoV-2 RNA copy number measurement and satisfied result was obtained by using our proposed RMP |
| Quantity | Amount-of-substance concentration (RNA copy number concentration) |
| From (0) | 15 |
| To (0) | 1000000 |
| Unit (0) | cp µL-1 |
| From (1) | 18 |
| To (1) | 12 |
| Unit (1) | % |
| Level of Confidence (%) | 95% |
| Peer Reviewed Publication | Highly accurate and sensitive diagnostic detection of SARS-CoV-2 by digital PCR, Dong L et al, Talanta 2020 Interlaboratory assessment of quantification of SARS-CoV-2 RNA by reverse transcription digital PCR, Niu C. et al, Analytical and Bioanalytical Chemistry 2021 413(29) 7195-7204 |
| Comments to be published via JCTLM DB | This reference measurement procedure (RMP) specifies the operating specifications for SARS-CoV-2 (open reading frame 1ab,ORF1ab and nucleocapsid protein, N and Envolpe, E gene) RNA copy number concentration by reverse transcription digital PCR (RT-dPCR) in in vitro RNA or RNA extracts from matrices in buffered solution. The approximate measurement interval is between 15 cp/μL to 106 cp/μL (equal to 100 cp/reaction to 106 cp/reaction). The upper limit of the linearity was 7.8 cp/partition tested by the N gene assay. Reactions containing a mean of 60 E, 66 N or 11 ORF1ab cp/reaction fulfilled the criterion for an Limit of quantification (LoQ) with a CV lower than 25%. The Limit of Blank (LoB) is determined to be 1.6, 1.6, and 0.8 cp/reaction for E, ORF1ab and N, respectively. The Limit of Detection (LoD) is determined to be 2 cp/reaction. As the RMP is restricted to purified RNA sample so the requirement for biosaftety is level 1. The performance of the extraction is not assured by RMP. Users should check isolated RNA samples for the presence of inhibitors which could interfere with RT-dPCR measurements. Users should review primer and probe sequences against prevalent SARS-CoV-2 strains at the time of application for any mismatches which would make the RMP unsuitable for quantification of SARS-CoV-2 gRNA targets. |
| Analyte Category Name | Nucleic acids |
| Non-CCQM Key Comparison Report | See Anal. Bioanal. Chem. 2021 413 7195 for national interlaboratory study |
| Measurement principle/technique | Digital PCR |