通过PCR-IS6110评估六种不同DNA提取方法用于检测结核分枝杆菌：初步研究
Evaluation of six different DNA extraction methods for detection of Mycobacterium tuberculosis by means of PCR-IS6110: preliminary study.
作者信息
De Almeida Isabela Neves, Da Silva Carvalho Wânia, Rossetti Maria Lúcia, Costa Elis Regina Dalla, De Miranda Silvana Spindola
机构信息
Research Group Coordinator FM/UFMG_REDE-TB, Belo Horizonte, Minas Gerais, Brazil.
出版信息
BMC Res Notes. 2013 Dec 28;6:561. doi: 10.1186/1756-0500-6-561.
BACKGROUND
Developments in molecular detection and strain differentiation of members of Mycobacterium tuberculosis complex have proved to be useful. The DNA extraction method influences the amplification efficiency, causing interference on the sensitivity and respective inhibitors. The aim of this study was to standardize a simple and fast DNA extraction method, providing DNA amplification by IS6110-PCR effectively free from undue interferences.
FINDINGS
The efficiency of the six different protocols tested in M. tuberculosis cultures has varied from 75% to 92.5%. This preliminary study evaluating the IS6110 PCR sensitivity and specificity was developed in DNA extracted from microscope slides, and achieved 100% of efficiency.
CONCLUSIONS
DNA extraction by Chelex + NP-40 method from both, cultures of M. tuberculosis and smear slides, resulted in good quantity of interference free DNA, especially in samples with low concentrations of genetic material; therefore, such technique may be used for the molecular diagnosis of tuberculosis.
背景
结核分枝杆菌复合群成员的分子检测和菌株鉴别技术已被证明是有用的。DNA提取方法会影响扩增效率，对灵敏度产生干扰并存在相应的抑制剂。本研究的目的是标准化一种简单快速的DNA提取方法，有效提供无不当干扰的IS6110-PCR DNA扩增。
研究结果
在结核分枝杆菌培养物中测试的六种不同方案的效率在75%至92.5%之间变化。这项评估IS6110 PCR灵敏度和特异性的初步研究是在从显微镜载玻片提取的DNA中进行的，效率达到了100%。
结论
采用Chelex + NP-40方法从结核分枝杆菌培养物和涂片载玻片中提取DNA，均能获得大量无干扰的DNA，尤其是在遗传物质浓度较低的样本中；因此，该技术可用于结核病的分子诊断。
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