噻唑蓝快速测定不同抗结核药物对卡介苗活性的影响

目的 利用黄色染料噻唑蓝(MTT) 可被活细胞消耗成紫色的甲月替结晶,溶解后可通过分光光度法进行定量测定的原理,快速检测体外培养卡介苗对不同浓度抗结核药物的耐药性,从而建立一种快速结核耐药测定方法?方法 用酶联免疫检测仪测定四种抗结核药物利福平,链霉素,异烟肼,对氨基水杨酸在不同浓度下对卡介苗活性的影响程度?结果 卡介苗总数与MTT的消耗量呈线性关系,死亡的卡介苗不能消耗MTT,不同浓度的抗结核药物对BCG 消耗MTT 的抑制作用不同,呈剂量_ 效应关系?药物浓度高低与培养菌落数呈线形关系?结论 该法快速,敏感,便宜,可在3 天内得到结果并可用肉眼观察,结果稳定,重复性好,可用于快速测定不同抗结核药物对卡介苗活性的影响,并极有可能用于临床快速检测结核杆菌的耐药情况?

Use of MTT for rapid detection of BCG viability influence of different anti_mycobacterial drugs

Objective To use the ability of viable cells to reduce 34,5dimethylthiazol2yi2,5diphenyltetrazolium bromide(MTT)to set up a test to detect 4 antimycobacterial drugs'influence on viability of invitro cultured BCG.Method To detect different concentration of 4 antimycobacterial drugs—Rifampin,Streptomycin,Isoniazid and Paminosalicylic acid's viability influent level in BCG.Result The assay shows a linear relationship between the number of vibable bacteria and the ability to reduce MTT.BCG exposed to antimycobacterial drugs showed concentrationdependent inhibition of the ability to reduce MTT.The inhibition paralleled the reduction in the number of colony forming units(CFU).Conclusion The assay can be visually read and requires less than 3 days to obtain susceptibility results.It is a cheap and rapid screening method for BCG vitality and be possible a cheap and rapid screening method for multidrugresistant M.tuberculosis.