Journal: Drug Testing and Analysis

Article Title: Improvement of EPO Transgene Detection From Polymeric Dried Blood Spots for Antidoping Application

doi: 10.1002/dta.70008

Figure Lengend Snippet: Comparison of EPO transgene detection from Tasso (T), VAMS (V), and Whatman(W) DBS collected with blood spiked at 1500 c/mL RM‐EPO. (A) Initial Testing procedure results using NMI ITP assay for qPCR. Cq values are indicated. NTC (no DNA in the PCR), PTC (160 copies of RM‐EPO in the PCR reaction). Peak of Melting and size of amplified products are shown for A and B samples. (B) Initial Testing procedure results using EPO Taqman assay for qPCR. Size of amplified products are shown for A and B samples. (C) Confirmation analysis results. Using NMI CP assay for qPCR. Cq values are indicated. NTC (no DNA in the PCR), PTC (160 copies of RM‐EPO in the PCR reaction). Size of amplified products are shown.

Article Snippet: EPO primers used for the initial testing procedure (ITP) were the previously validated Taqman 20× Gene Expression Assay for EPO (Hs01071097_m1) (ThermoFisher Scientific) or NMI specific primers for the initial testing procedure (NMI ITP 10×) [ , ] both targeting the exon 3–exon 4 junction from the human EPO gene but with different sizes of amplified products (NMI ITP assay: 64 base pairs (bp) for RM‐EPO; EPO Taqman assay: 123 bp for RM‐EPO).

Techniques: Comparison, Amplification, TaqMan Assay, Ceruloplasmin Assay

Journal: Drug Testing and Analysis

Article Title: Improvement of EPO Transgene Detection From Polymeric Dried Blood Spots for Antidoping Application

doi: 10.1002/dta.70008

Figure Lengend Snippet: Validation of RM‐EPO detection from TASSO DBS. A. Selectivity, sensitivity (Limit of detection) and reproducibility of detection are shown. PCR was performed using Taqman EPO assay on DNA extracted from DBS TASSO M‐20 loaded with 10 different blood samples spiked at 0–1500–2500–5000 c/mL RM‐EPO. Cq values are indicated. Examples of size of amplified products are shown for bloods spiked at 5000 c/mL. Reproducibility experiments were performed after DNA extraction from the second and third spot of the TASSO device. B. CP performed on the fourth spot of TASSO M‐20 (“B analysis”). PCR was performed using NMI CP on DNA extracted from the fourth spot (last spot) of DBS TASSO M‐20 loaded with blood samples spiked at 5000 c/mL RM‐EPO. Cq values are indicated. Examples of size of amplified products are shown.

Article Snippet: EPO primers used for the initial testing procedure (ITP) were the previously validated Taqman 20× Gene Expression Assay for EPO (Hs01071097_m1) (ThermoFisher Scientific) or NMI specific primers for the initial testing procedure (NMI ITP 10×) [ , ] both targeting the exon 3–exon 4 junction from the human EPO gene but with different sizes of amplified products (NMI ITP assay: 64 base pairs (bp) for RM‐EPO; EPO Taqman assay: 123 bp for RM‐EPO).

Techniques: Biomarker Discovery, Amplification, DNA Extraction