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2,3-双(2-甲氧基-4-硝基-5-磺基苯基)-2H-四唑-5-甲酰苯胺(XTT)试验作为一种快速比色法,用于测定液体培养基中临床结核分枝杆菌分离株的抗生素敏感性。

The 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay as rapid colorimetric method for determination of antibiotic susceptibility of clinical Mycobacterium tuberculosis isolates in liquid medium.

作者信息

De Logu Alessandro, Borgna Rita, Uda Patrizia, Sanna Adriana, Pellerano Maria Luisa, Saddi Barbara

机构信息

Division of Medical Microbiology, Department of Biomedical Sciences and Technologies, University of Cagliari, Cagliari, Italy.

出版信息

Clin Lab. 2003;49(7-8):357-65.

Abstract

We investigated the usefulness of a colorimetric method based on the reduction of a tetrazolium salt (XTT) for the susceptibility testing of clinical isolates of Mycobacterium tuberculosis to isoniazid, rifampin, rifabutin, ethambutol hydrochloride, ethionamide and streptomycin. The isolates and the ATCC reference strains reported as susceptible according to the agar dilution method approved by the National Committee for Clinical Laboratory Standards were found to be susceptible by the XTT colorimetric assay after times of incubation ranging between three days for rifampin and rifabutin to eight days for isoniazid. In comparison with other colorimetric methods reviewed in this article, the proposed assay is suitable for determining the susceptibility or resistance to most antituberculous drugs and, as a consequence of the water-solubility of the formazan yielded by reduction of XTT, additional steps such as the addition of extraction buffer and further incubation before the spectrophotometric analysis are not needed. The XTT reduction assay is an inexpensive, rapid and reliable screening method for the detection of susceptible, resistant and multidrug-resistant strains of M. tuberculosis and is an alternative to the costly performance of molecular or radiometric methods.

摘要

我们研究了一种基于四氮唑盐(XTT)还原的比色法在检测结核分枝杆菌临床分离株对异烟肼、利福平、利福布汀、盐酸乙胺丁醇、乙硫异烟胺和链霉素敏感性方面的实用性。根据美国国家临床实验室标准委员会批准的琼脂稀释法报告为敏感的分离株和ATCC参考菌株,经XTT比色法检测,孵育时间从利福平和利福布汀的3天到异烟肼的8天不等,结果均显示敏感。与本文中综述的其他比色法相比,该方法适用于确定对大多数抗结核药物的敏感性或耐药性,并且由于XTT还原产生的甲臜具有水溶性,无需在分光光度分析前添加提取缓冲液和进一步孵育等额外步骤。XTT还原试验是一种用于检测结核分枝杆菌敏感、耐药和多重耐药菌株的廉价、快速且可靠的筛选方法,是昂贵的分子或放射性方法的替代方法。

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