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1.发明公开METHOD FOR THE MICROBIOLOGICAL DETERMINATION OF TRACES OF ANTIBIOTICS IN LOW VOLUME BIOLOGICAL SAMPLES 审中-公开方法抗生素的痕量在低体积生物样品微生物学测定
公开(公告)号：EP3085790A1
公开(公告)日：2016-10-26
申请号：EP14872796.9
申请日：2014-12-11
发明人： SANHUEZA CHAVEZ, Loreto , WILKENS ANWANDTER, Marcela Andrea , LAURIDO FUENZALIDA, Claudio Aurelio
CPC分类号： C12Q1/18 , G01N2333/31
摘要： A microbiological method for determining traces of antibiotics in low volume biological samples, which comprises determining the minimum inhibitory concentration (MIC) of an antibiotic selected from a group consisting of minocycline, ciprofloxacin, kanamycin, tetracycline and oxacillin against Staphylococcus aureus. The bacteria are grown overnight in Mueller-Hinton broth (MHB), said cultures subsequently being diluted to 0.5 McFarland standard (1.5 x 108 cells/mL). Said antibiotics are then added to 96-well plates to a final volume of 200 µL and incubated at 37°C for 18 hours. Absorbency is determined and the results are expressed as the percentage of inhibition relative to the control containing the biological sample, selected in turn from urine, blood, synovial fluid, eye aqueous humour or cerebrospinal fluid (CSF), and the bacterial culture. The curve representing the percentage of bacterial growth inhibition versus concentration of antibiotic is obtained, and fitted by "Four Parameter Logistic Curve (4PL)" analysis, yielding the antibiotic concentration values in the biological sample. The method allows to determine amounts of antibiotic in the biological sample even at ng/mL level.
摘要翻译： 对于低体积的生物样品中的抗生素确定性采矿迹线的微生物学方法，该方法包括确定性采矿从包括米诺环素，环丙沙星，卡那霉素，四环素和苯唑西林对金黄色葡萄球菌的选择的抗生素的最小抑制浓度（MIC）。 该细菌在Mueller Hinton肉汤（MHB）中生长过夜，说培养随后稀释至0.5麦氏标准（1.5×108细胞/ mL）。 然后，所述抗生素加入到96孔板中，以200μl的最终体积，并在37℃下温育18小时。 吸收能力被确定性开采并且将结果表示为相对于抑制的百分比包含所述生物样品，依次从尿，血液，滑液，眼睛wässrige幽默或脑脊髓液（CSF）中选择的控制，和细菌培养。 代表的细菌生长抑制与抗生素的浓度的百分比的曲线被获得，并且通过“四参数逻辑曲线（4PL）”分析嵌合，在生物样品中得到的抗菌素的浓度值。 该方法允许甚至在毫微克/毫升的水平的生物样品中的抗生素的确定性矿量。
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公开(公告)号：EP3085790B1
公开(公告)日：2018-08-29
申请号：EP14872796.9
申请日：2014-12-11
发明人： SANHUEZA CHAVEZ, Loreto , WILKENS ANWANDTER, Marcela Andrea , LAURIDO FUENZALIDA, Claudio Aurelio
CPC分类号： C12Q1/18 , G01N2333/31
摘要： A microbiological method for determining traces of antibiotics in low volume biological samples, which comprises determining the minimum inhibitory concentration (MIC) of an antibiotic selected from a group consisting of minocycline, ciprofloxacin, kanamycin, tetracycline and oxacillin against Staphylococcus aureus. The bacteria are grown overnight in Mueller-Hinton broth (MHB), said cultures subsequently being diluted to 0.5 McFarland standard (1.5 x 108 cells/mL). Said antibiotics are then added to 96-well plates to a final volume of 200 µL and incubated at 37°C for 18 hours. Absorbency is determined and the results are expressed as the percentage of inhibition relative to the control containing the biological sample, selected in turn from urine, blood, synovial fluid, eye aqueous humour or cerebrospinal fluid (CSF), and the bacterial culture. The curve representing the percentage of bacterial growth inhibition versus concentration of antibiotic is obtained, and fitted by "Four Parameter Logistic Curve (4PL)" analysis, yielding the antibiotic concentration values in the biological sample. The method allows to determine amounts of antibiotic in the biological sample even at ng/mL level.
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