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赛马非同位素免疫分析药物检测:其在赛前和赛后检测、药物定量及人体药物检测中的应用综述

Non-isotopic immunoassay drug tests in racing horses: a review of their application to pre- and post-race testing, drug quantitation, and human drug testing.

作者信息

Tobin T, Watt D S, Kwiatkowski S, Tai H H, Blake J W, McDonald J, Prange C A, Wie S

机构信息

Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington 40546-0099.

出版信息

Res Commun Chem Pathol Pharmacol. 1988 Dec;62(3):371-95.

PMID:3065859
Abstract

We have introduced large scale non-isotopic immunoassay testing into pre- and post-race drug testing in racehorses. The technologies utilized are Particle Concentration Fluorescence Immuno Assay (PCFIA) and the one-step Enzyme Linked Immuno Sorbent Assay (ELISA). These technologies are rapid, inexpensive, and highly effective. On introduction into post-race testing in the Western United States, these ELISA tests exposed several previously undetected patterns of drug abuse. The drugs detected were buprenorphine, oxymorphone, mazindol, sufentanil and cocaine. This led to the suspension of a large number of trainers and exposed the high false negative rate of thin layer chromatography (TLC) based testing. More recently, we have introduced both PCFIA and ELISA assays into pre- and post-race testing in Illinois. Within days, our pre-race PCFIA tests detected signs of acepromazine abuse. Directed searches of post-race urines from these horses showed evidence for acepromazine metabolites in the urine of these horses. Examination of frozen samples from associated horses yielded about 70 ELISA "positives" for acepromazine. To date, about 25 of these ELISA "positives" have been confirmed by mass spectrometry. We have also raised antibodies to phenylbutazone and furosemide to enable rapid and inexpensive quantitation of these permitted medications. Furosemide is a particular problem since its use requires a pre-race detention barn. For furosemide, we have developed a regulatory schedule based on our immunoassay test that allows elimination of the detention barn. For phenylbutazone, we have developed a similar immunoassay that allows rapid and inexpensive quantitation of this drug in blood. To enable racing authorities to test jockeys and other racetrack personnel, we have adapted PCFIA technology to human drug testing, and a full range of very sensitive tests for human drugs of abuse is available. These immunoassays are sufficiently sensitive to control abuse of the most potent drugs available to horsemen. In principle, an immunoassay can be raised to any drug within about six months, and made available worldwide at competitive rates. It appears clear that these non-isotopic immunoassays provide racing with the only technological basis that is sufficiently sensitive to detect the most potent abused drugs pre- and post-race, and has the flexibility to be readily adaptable to different drugs. Because of the high false negative rate generated by TLC, credible pre- and post-race testing programs cannot be based on TLC alone.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

我们已将大规模非同位素免疫分析检测引入赛马赛前和赛后药物检测中。所采用的技术是颗粒浓度荧光免疫分析(PCFIA)和一步式酶联免疫吸附测定(ELISA)。这些技术快速、廉价且高效。在美国西部引入赛后检测时,这些ELISA检测发现了几种此前未被检测到的药物滥用模式。检测出的药物有丁丙诺啡、羟吗啡酮、马吲哚、舒芬太尼和可卡因。这导致大量驯马师被停职,并暴露了基于薄层色谱(TLC)检测的高假阴性率。最近,我们已将PCFIA和ELISA检测引入伊利诺伊州的赛前和赛后检测。几天内,我们的赛前PCFIA检测就发现了乙酰丙嗪滥用的迹象。对这些马匹赛后尿液的定向检测显示这些马的尿液中有乙酰丙嗪代谢物的证据。对相关马匹的冷冻样本检测产生了约70个乙酰丙嗪的ELISA“阳性”结果。迄今为止,这些ELISA“阳性”结果中约有25个已通过质谱法得到确认。我们还制备了针对保泰松和呋塞米的抗体,以便对这些许可使用的药物进行快速且廉价的定量检测。呋塞米是个特别的问题,因为使用它需要赛前隔离厩。对于呋塞米,我们根据免疫分析检测制定了一项监管计划,从而可以不再需要隔离厩。对于保泰松,我们开发了一种类似的免疫分析方法,能够对血液中的这种药物进行快速且廉价的定量检测。为使赛马管理机构能够对骑师和其他赛道工作人员进行检测,我们已将PCFIA技术应用于人体药物检测,并且有一整套针对人体滥用药物的非常灵敏的检测方法。这些免疫分析方法灵敏度足够高,能够控制骑手可获取的最有效药物的滥用情况。原则上,针对任何药物的免疫分析方法大约在六个月内即可研发出来,并能以有竞争力的价格在全球范围内提供。显然,这些非同位素免疫分析为赛马运动提供了唯一一种技术基础,它灵敏度足够高,能够在赛前和赛后检测出最有效的滥用药物,并且具有易于适应不同药物的灵活性。由于TLC产生的高假阴性率,可靠的赛前和赛后检测项目不能仅基于TLC。(摘要截选至400字)

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