Evidence mapPaperPMID 42065780Full record

ArticleAnalytical and bioanalytical chemistry2026

Direct LC-HRMS/MS characterisation and RT-qPCR detection for potential mRNA-based doping agent.

Bruce P-N Yuen, Kin-Sing Wong, Venus Y-C Lin, Hiu Wing Cheung, Emmie N-M Ho, Wing-Tak Wong

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In one paragraph

Article in Analytical and bioanalytical chemistry, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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0citing papers in PubMed
field-weighted citation impact
1 · What the graph read from it

What it found

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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

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Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

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0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

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PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors.

Bruce P-N YuenDepartment of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Kowloon, Hong Kong, China.
Kin-Sing WongRacing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N.T., Hong Kong, China.
Venus Y-C LinRacing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N.T., Hong Kong, China.
Hiu Wing CheungRacing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N.T., Hong Kong, China.
Emmie N-M HoRacing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N.T., Hong Kong, China. emmie.nm.ho@hkjc.org.hk.
Wing-Tak WongDepartment of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Kowloon, Hong Kong, China.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Messenger ribonucleic acid (mRNA) therapeutics offer unprecedented potential for in vivo protein expression but raise emerging risks of misuse for gene doping. We investigated an online-available product purported to be codon-optimised, 5-methoxyuridine-substituted human erythropoietin (EPO) mRNA and developed a corresponding detection assay for equine anti-doping. Product identity was verified using a mass spectrometry (MS)-based bottom-up workflow comprising RNase 4 digestion, liquid chromatography-high-resolution tandem MS analysis, and automated sequence mapping against an mRNA database of potential doping genes. This workflow enabled direct detection of base modifications and achieved mean sequence coverage above 74%, verifying the mRNA sequence and its chemical composition. Method optimisation shows that partial RNase 4 digestion with a 10-min incubation time produced more consistent coverage by generating longer, informative oligoribonucleotides. The enhanced consistency would be advantageous for single-analysis scenarios typical of doping investigations. Building on the verified product identity, we developed a corresponding reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay for equine plasma. We explored the simple use of naked mRNA for matrix spiking as a reference material for doping control analysis. The method was validated with a limit of detection at 1250 copies/mL of EPO mRNA in equine plasma. In addition to EPO mRNA, the combined MS and RT-qPCR approach provides a practical framework that can be extended for surveillance of emerging mRNA-based agents in sport.

Indexed as

Doping in SportsErythropoietinHorsesLiquid Chromatography-Mass SpectrometryReverse Transcriptase Polymerase Chain ReactionRNA, MessengerAnimalsTandem Mass SpectrometryErythropoietinRNA, MessengerEquine sportsGene doping detectionLC–MSMappingmRNA therapeuticsRT–qPCR

Identifiers

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Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.