高效液相色谱与荧光法测定细菌染色体碱基组成的比较 |
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引用本文: | 许化溪,王胜军,黄锡全. 高效液相色谱与荧光法测定细菌染色体碱基组成的比较[J]. 中华检验医学杂志, 2000, 23(4): 237-240 |
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作者姓名: | 许化溪 王胜军 黄锡全 |
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作者单位: | [1]江苏省镇江医学院检验医学系 [2]日本岐阜大学医学部微生物学教室 |
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摘 要: | 目的 比较连续荧光检测法(荧光法)与高效液相色谱(HPLC)法测定细菌染色体鸟嘌呤胞嘧啶百分比(GC%)的稳定性、准确性和适应性。方法 应用双股DNA结合染料和能够快速速升降温度的连续荧光检测仪、测定9株细菌染色体的变性温度℃值,并根据相应的公式算出GC%:C HPLC法直接检测细菌染色体的GC%。结果 DNA纯度对荧光法测得的GC%结果无显著影响,但影响HPLC的测定结果,荧光法3次检测获得的9
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关 键 词: | 细菌染色体 高效液相色谱法 连续荧光检测仪 |
修稿时间: | 1999-08-09 |
Light cycler and high pressure liquid chromatography in determing the guanine plus cytosine content of bacterial chromosome DNA |
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Abstract: | Objective To compare stability, accuracy and suitability of Light Gycler and high prescure liquid chromatography in detecting mole percent guanine plus cytosine content (GC%).Methods Light Cycler method, was used following the melting curves and the values of Tm were obtained from the computer screen of Light Cycler, to calculate the GC% according to the formula: GC%x=GC%r - 8.567 (Tmx Tmr) 0.066 (Tmx 2 Tmr 2). The GC% of bacterial chromosome DNA was obtained directly by HPLC within 30 min after the samples were loaded. Results By Light Cycler we detected the values of the Tm from 24 samples obtained within 10 min. The purity of DNA was not visiblely affect the result. The GC% from HPLC method was stable and accurate, and it was more similar to the published values. But the procedure for further purification of DNA should be included.Conclusion The Light Cycler is approprate for clinical diagnosis of bacteria because. The HPLC method is accurate and reliable for bacterial classification. |
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Keywords: | Bacterial stain High performance liquid chromatography Light Cycler |
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