[Comparative analysis of RT-qPCR based methodologies for microRNA detection.] 2018

E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
N.N. Petrov Research Institute of Oncology, 197758, Saint Petersburg, Russia.

Many pathological states are accompanied by characteristic changes in the cellular profile of microRNAs - small molecules that regulate gene expression at the posttranscriptional level. This allows us to consider miRNA as a promising class of biological markers. In the work, a direct comparison of three RT-qPCR methodologies (s-Loop, u-Elong and 2-Tail) for miRNA analysis was performed. A synthetic miRNA-451 analog was used to determine the efficiency of miRNA molecule detection and analysis of the miRNA-29b, miRNA-375 and miRNA-451 profiles in OAW42 and HT29 cell lines was carried out. By the methods of 2-Tail and s-Loop, seven different miRNA were also analyzed in 13 clinical specimens. The results of the study show that in the 2-Tail and s-Loop approaches, RT-qPCR demonstrated high reproducibility in results of miRNA analysis, and a linear dependence of the mimic миРНК-451detection efficiency in the range of 107 to 103 molecules per reaction was registered. On a number of significant criteria, the two technologies turned out to be relatively equivalent, i.e. any of them can be used as a basis for the method of clinical diagnostics.

UI MeSH Term Description Entries
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D015203 Reproducibility of Results The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results. Reliability and Validity,Reliability of Result,Reproducibility Of Result,Reproducibility of Finding,Validity of Result,Validity of Results,Face Validity,Reliability (Epidemiology),Reliability of Results,Reproducibility of Findings,Test-Retest Reliability,Validity (Epidemiology),Finding Reproducibilities,Finding Reproducibility,Of Result, Reproducibility,Of Results, Reproducibility,Reliabilities, Test-Retest,Reliability, Test-Retest,Result Reliabilities,Result Reliability,Result Validities,Result Validity,Result, Reproducibility Of,Results, Reproducibility Of,Test Retest Reliability,Validity and Reliability,Validity, Face
D015415 Biomarkers Measurable and quantifiable biological parameters (e.g., specific enzyme concentration, specific hormone concentration, specific gene phenotype distribution in a population, presence of biological substances) which serve as indices for health- and physiology-related assessments, such as disease risk, psychiatric disorders, ENVIRONMENTAL EXPOSURE and its effects, disease diagnosis; METABOLIC PROCESSES; SUBSTANCE ABUSE; PREGNANCY; cell line development; EPIDEMIOLOGIC STUDIES; etc. Biochemical Markers,Biological Markers,Biomarker,Clinical Markers,Immunologic Markers,Laboratory Markers,Markers, Biochemical,Markers, Biological,Markers, Clinical,Markers, Immunologic,Markers, Laboratory,Markers, Serum,Markers, Surrogate,Markers, Viral,Serum Markers,Surrogate Markers,Viral Markers,Biochemical Marker,Biologic Marker,Biologic Markers,Clinical Marker,Immune Marker,Immune Markers,Immunologic Marker,Laboratory Marker,Marker, Biochemical,Marker, Biological,Marker, Clinical,Marker, Immunologic,Marker, Laboratory,Marker, Serum,Marker, Surrogate,Serum Marker,Surrogate End Point,Surrogate End Points,Surrogate Endpoint,Surrogate Endpoints,Surrogate Marker,Viral Marker,Biological Marker,End Point, Surrogate,End Points, Surrogate,Endpoint, Surrogate,Endpoints, Surrogate,Marker, Biologic,Marker, Immune,Marker, Viral,Markers, Biologic,Markers, Immune
D060888 Real-Time Polymerase Chain Reaction Methods used for detecting the amplified DNA products from the polymerase chain reaction as they accumulate instead of at the end of the reaction. Kinetic Polymerase Chain Reaction,Quantitative Real-Time PCR,Quantitative Real-Time Polymerase Chain Reaction,Real-Time PCR,PCR, Quantitative Real-Time,PCR, Real-Time,PCRs, Quantitative Real-Time,PCRs, Real-Time,Quantitative Real Time PCR,Quantitative Real Time Polymerase Chain Reaction,Quantitative Real-Time PCRs,Real Time PCR,Real Time Polymerase Chain Reaction,Real-Time PCR, Quantitative,Real-Time PCRs,Real-Time PCRs, Quantitative
D019073 HT29 Cells Human colonic ADENOCARCINOMA cells that are able to express differentiation features characteristic of mature intestinal cells such as the GOBLET CELLS. HT-29 Cells,Cell, HT-29,Cell, HT29,Cells, HT-29,Cells, HT29,HT 29 Cells,HT-29 Cell,HT29 Cell
D035683 MicroRNAs Small double-stranded, non-protein coding RNAs, 21-25 nucleotides in length generated from single-stranded microRNA gene transcripts by the same RIBONUCLEASE III, Dicer, that produces small interfering RNAs (RNA, SMALL INTERFERING). They become part of the RNA-INDUCED SILENCING COMPLEX and repress the translation (TRANSLATION, GENETIC) of target RNA by binding to homologous 3'UTR region as an imperfect match. The small temporal RNAs (stRNAs), let-7 and lin-4, from C. elegans, are the first 2 miRNAs discovered, and are from a class of miRNAs involved in developmental timing. RNA, Small Temporal,Small Temporal RNA,miRNA,stRNA,Micro RNA,MicroRNA,Primary MicroRNA,Primary miRNA,miRNAs,pre-miRNA,pri-miRNA,MicroRNA, Primary,RNA, Micro,Temporal RNA, Small,miRNA, Primary,pre miRNA,pri miRNA

Related Publications

E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
January 2020, Methods in molecular biology (Clifton, N.J.),
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
April 2010, Methods (San Diego, Calif.),
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
March 2020, Molecules (Basel, Switzerland),
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
October 2023, Letters in applied microbiology,
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
January 2017, Methods in molecular biology (Clifton, N.J.),
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
February 2019, Chemical science,
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
January 2012, Methods in molecular biology (Clifton, N.J.),
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
November 2017, BMC research notes,
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
January 2017, Biosensors & bioelectronics,
E A Korobkina, and M S Knyazeva, and Yu V Kil, and S E Titov, and A V Malek
November 2022, Diagnostics (Basel, Switzerland),
Copied contents to your clipboard!