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首页> 美国卫生研究院文献>Advanced Pharmaceutical Bulletin >Study of the Efficacy of Real Time-PCR Method for Amikacin Determination Using Microbial Assay
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Study of the Efficacy of Real Time-PCR Method for Amikacin Determination Using Microbial Assay

机译：实时荧光定量PCR方法测定阿米卡星微生物含量的功效研究

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>Purpose: Microbial assay is used to determine the potency of antibiotics and vitamins. In spite of its advantages like simplicity and easiness, and to reveal the slight changes in the molecules, the microbial assay suffers from significant limitations; these methods are of lower specificity, accuracy and sensitivity. The objective of the present study is to evaluate the efficacy of real time-PCR technique in comparison with turbidimetric method for microbial assay of amikacin.>Methods: Microbial determination of amikacin by turbidimetric method was performed according to USP. Also amikacin concentrations were determined by microbial assay using taq-man quantitative PCR method. Standard curves in different concentration for both methods were plotted and method validation parameters of linearity, precision and accuracy were calculated using statistical procedures.>Results: The RT-PCR method was linear in the wider concentration range (5.12 – 38.08 for RT-PCR versus 8.00 – 30.47 for turbidimetric method) with a better correlation coefficient (0.976 for RT-PCR versus 0.958 for turbidimetric method). RT-PCR method with LOQ of 5.12 ng/ml was more sensitive than turbidimetric method with LOQ of 8.00 ng/ml and the former could detect and quantify low concentrations of amikacin. The results of accuracy and precision evaluation showed that the RT-PCR method was accurate and precise in all of the tested concentration.>Conclusion: The RT-PCR method described here provided an accurate and precise technique for measurement of amikacin potency and it can be a candidate for microbial determination of the antibiotics with the same test organism.
机译：>目的：微生物检测可用于确定抗生素和维生素的效力。尽管它具有简单，易用等优点，并且可以揭示分子中的细微变化，但微生物检测仍存在明显的局限性。这些方法的特异性，准确性和敏感性较低。本研究的目的是评估实时PCR技术与比浊法比较阿米卡星的微生物含量的有效性。>方法：根据USP进行比浊法对阿米卡星的微生物测定。还使用taq-man定量PCR方法通过微生物测定法确定了丁胺卡那霉素的浓度。绘制了两种方法在不同浓度下的标准曲线，并使用统计程序计算了线性，精密度和准确性的方法验证参数。>结果： RT-PCR方法在较宽的浓度范围内呈线性（5.12 – RT-PCR为38.08，比浊法为8.00 – 30.47）具有更好的相关系数（RT-PCR为0.976，比浊法为0.958）。 LO-为5.12 ng / ml的RT-PCR方法比浊度为8.00 ng / ml的比浊法更灵敏，前者可以检测和定量低浓度的阿米卡星。准确度和精密度评估结果表明，RT-PCR方法在所有测试浓度下都是准确和精确的。>结论：此处介绍的RT-PCR方法提供了一种精确，准确的技术来测量丁胺卡那霉素的效力，可以与同一受试微生物一起对抗生素进行微生物测定。

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