Detection of intra-articular gene therapy in horses using quantitative real time PCR in synovial fluid and plasma.

Drug Testing and Analysis
Joanne HaughanMary A Robinson

Abstract

Gene therapy promotes the expression of missing or defective genes and can be curative following administration of a single dose. Gene therapy is prohibited in equine athletes by regulatory bodies due to the high potential for abuse and novel analytical methods are needed for detection. The goal of this study was to detect the administration of an experimental gene therapy: a recombinant adeno-associated viral vector (rAAV) carrying a transgene for the anti-inflammatory cytokine IL-10 (rAAV-IL10). Twelve horses were randomly assigned to receive an intra-articular injection of rAAV-IL10 or phosphate buffered saline (vehicle) into a middle carpal joint. Plasma and synovial fluid were collected on days 0, 1, 2, 4, 7, 14, 28, 56, and 84. Primer pairs were designed to detect two unique regions of rAAV. Using quantitative real time PCR, both sets of primers detected rAAV for 14-28 days in joints and up to 4 days in plasma, in all six treated horses. In synovial fluid, rAAV was detected on day 56 in 4/6 horses by both primer sets, and on day 84 in 1/6 horses by one primer set. In plasma, rAAV was detected for 7 days in 5/6 horses, 14 days in 2/6 horses, and 28 days in 1/6 horses by one primer set, and was detected for up to 14 days in...Continue Reading

References

Feb 7, 2001·Journal of Clinical Microbiology·W A Al-Soud, P Rådström
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Aug 27, 2009·Proceedings of the National Academy of Sciences of the United States of America·Mark L BrantlyTerence R Flotte
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Oct 16, 2015·Nature·Luigi Naldini
Mar 30, 2017·Drug Testing and Analysis·Tessa WilkinNatasha Hamilton
Jun 5, 2017·Medicine and Sport Science·Elmo W I Neuberger, Perikles Simon
Jul 3, 2017·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Michael F NasoWilliam R Strohl
Jul 14, 2017·Biomolecular Detection and Quantification·Amin ForootanMikael Kubista
Mar 27, 2018·The Journal of Gene Medicine·Samantha L GinnMohammad R Abedi

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Citations

Apr 30, 2021·Analytical Chemistry·Teruaki TozakiShunichi Nagata
Aug 8, 2021·Scientific Reports·Teruaki TozakiShun-Ichi Nagata
Aug 20, 2021·Drug Testing and Analysis·Zibin JiangMary A Robinson
Oct 6, 2021·Drug Testing and Analysis·Teruaki TozakiShun-Ichi Nagata

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