qPCR and qRT-PCR analysis: Regulatory points to consider when conducting biodistribution and vector shedding studies.
| Autor: | Ma H; Northern Biomolecular Services, 4717 Campus Drive, Kalamazoo, MI 49008, USA., Bell KN; Northern Biomolecular Services, 4717 Campus Drive, Kalamazoo, MI 49008, USA., Loker RN; Northern Biomolecular Services, 4717 Campus Drive, Kalamazoo, MI 49008, USA. |
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| Jazyk: | angličtina |
| Zdroj: | Molecular therapy. Methods & clinical development [Mol Ther Methods Clin Dev] 2020 Nov 17; Vol. 20, pp. 152-168. Date of Electronic Publication: 2020 Nov 17 (Print Publication: 2021). |
| DOI: | 10.1016/j.omtm.2020.11.007 |
| Abstrakt: | Gene and cell therapy fields have experienced remarkable growth during the past decade. Demands for preclinical and clinical safety assessments of these cell and gene therapy test articles (TAs) have effectively increased the necessity for regulated biodistribution, vector shedding, gene expression, and/or pharmacokinetics bioanalysis studies. Guidance documents issued from numerous international regulatory authorities recommend the use of quantitative polymerase chain reaction (qPCR) and/or quantitative reverse transcriptase PCR (qRT-PCR) assays due to their highly sensitive and robust target-specific detection. However, only preclinical biodistribution assay sensitivity is specified in these documents. Criteria such as accuracy, precision, and repeatability are not yet defined. This guidance void has resulted in several conflicting institutional interpretations of essential parameters necessary for the development and validation of robust assays to support safety assessments of gene and cell therapy TAs. There is an urgent need for an ongoing discussion among bioanalytical scientists in this field to generate a "best practice" consensus around preclinical and clinical qPCR/qRT-PCR assay design. With regard to this need, we offer critical points to consider when developing, validating, running sample analysis, and reporting qPCR/qRT-PCR assays. Competing Interests: All authors are employees of Northern Biomolecular Services, and may own respective stock of the company. The authors declare no additional competing interests. (© 2020 The Author(s).) |
| Databáze: | MEDLINE |
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