Development and evaluation of armored RNA-based standards for quantification of BCR-ABL1p210/p190 fusion gene transcripts.
Background: Standards play an important role in detection of the BCR-ABL1 fusion gene (FG) transcript. However, the standards widely used in laboratories are mainly based on plasmids or cDNA, which cannot accurately reflect the process of RNA extraction and cDNA synthesis. Therefore, we aimed to dev...
| Published in: | Journal of Clinical Laboratory Analysis Vol. 32; no. 9 |
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| Main Authors: | , , , , , |
| Format: | pictorial research tables/charts Journal Article |
| Published: |
Wiley-Blackwell
Nov2018
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| Online Access: | View this record in EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=132916184&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 132916184 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 08878013 OCK jtl: Journal of Clinical Laboratory Analysis issn: 08878013 maglogo: Y pubinfo: dt: Nov2018 vid: 32 iid: 9 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 132916184 132916184 NLM29959790 132916184 10.1002/jcla.22612 NLM29959790 132916184 ppct: 1 formats: tig: atl: Development and evaluation of armored RNA-based standards for quantification of BCR-ABL1p210/p190 fusion gene transcripts. aug: au: Fu, Yu Zhang, Rui Wu, Qisheng Zhang, Jiawei Bao, Lihua Li, Jinming affil: National Center for Clinical Laboratories, Beijing Hospital, National Center of Gerontology, Beijing China sug: subj: Proteins Analysis Polymerase Chain Reaction Methods RNA Analysis Proteins Statistics Reference Values Funding Source Human ab: Background: Standards play an important role in detection of the BCR-ABL1 fusion gene (FG) transcript. However, the standards widely used in laboratories are mainly based on plasmids or cDNA, which cannot accurately reflect the process of RNA extraction and cDNA synthesis. Therefore, we aimed to develop armored RNA-based standards for p210 and p190 BCR-ABL1FG transcripts' quantification.Methods: Using overlapping polymerase chain reaction (PCR) technology, we first linked a segment of the p210 or p190 BCR-ABL1FG transcript with four control genes (CGs; ABL1, BCR, GUSB, and B2M) to form p210FG-CG and p190FG-CG. Subsequently, using armored RNA technology, we prepared p210FG-CG- and p190FG-CG-armored RNAs and the p210FG-CG and p190FG-CG standards, the values of which were assigned by digital PCR (dPCR).Results: The p210FG-CG and p190FG-CG standards were stable and homogeneous, and were significantly linear with r2 > 0.98. A field trial including 52 laboratories across China showed that the coefficient of variation (CV%) of BCR-ABL1 values among samples was in the range of 58.6%-129.6% for p210 samples and 73.2%-194.0% for p190 samples when using local standards. By contrast, when using the p210FG-CG and p190FG-CG standards, the CV% of BCR-ABL1 values was decreased to 35.6%-124.9% and 36.6%-170.6% for p210 and p190 samples, respectively. In addition, 33.3% (3/9) of the p210 and p190 samples had CV% values <50.0%, whereas 44.4% (4/9) and 77.8% (7/9) of the samples had lower CV% values when using the p210FG-CG and p190FG-CG standards.Conclusion: The overall variability of detection of BCR-ABL1 transcripts decreased significantly when using the p210FG-CG or p190FG-CG standards, especially the p190FG-CG standard. pubtype: Academic Journal doctype: pictorial research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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