Isothermal DNA amplification coupled to Au-nanoprobes for detection of mutations associated to Rifampicin resistance in Mycobacterium tuberculosis.
Isothermal DNA amplification coupled to Au-nanoprobes for detection of mutations associated to Rifampicin resistance in Mycobacterium tuberculosis.
复制标题
DOI：
10.1186/1477-3155-11-38
复制
发表时间：
2013-11-25
影响因子：
10.2
通讯作者：
Baptista PV
中科院分区：
文献类型：
--
作者：
Veigas B;Pedrosa P;Couto I;Viveiros M;Baptista PV
2011年，结核病新增病例达870万例，仍是人类主要传染病之一。造成负担的是耐多药结核病(MDRTB)的增加，以及给治疗和公共卫生控制方案带来的困难，特别是在发展中国家。对一线抗生素利福平(RIF)的耐药性通常与结核分枝杆菌(Mtb)rpoB基因点突变有关，该基因的检测被认为是MDRTB最好的早期分子预测指标。用硫醇修饰的寡核苷酸功能化的金纳米颗粒(金纳米探针)有可能为与抗生素耐药相关的Mtb和单碱基改变，即与RIF耐药相关的rpoB基因的改变提供一种快速而灵敏的检测方法。我们开发了一种基于样品DNA(LAMP)与特定Au-纳米探针的等温扩增的策略，能够识别MTBC的成员并识别rpoB基因内的特定突变。LAMP和Au纳米探针检测的结合使得能够检测MTBC成员并识别与RIF耐药相关的突变。对12份生物样品进行了检测，特异性和敏感性均为100%。越来越多的人需要简单、快速和廉价的方法来鉴定结核分枝杆菌的分子，并检测适合在需要时使用的与耐药性相关的分子标签。在这里，我们描述了这样一种方法，即有可能在偏远环境中获得结核病的分子诊断。
Tuberculosis accounted for 8.7 million new cases in 2011 and continues to be one of the leading human infectious diseases. Burdensome is the increasing rate of multi-drug resistant tuberculosis (MDRTB) and the difficulties created for treatment and public health control programs, especially in developing countries. Resistance to rifampicin (RIF), a first line antibiotic, is commonly associated with point mutations within the rpoB gene of Mycobacterium tuberculosis (Mtb) whose detection is considered the best early molecular predictor for MDRTB. Gold nanoparticles functionalized with thiol-modified oligonucleotides (Au-nanoprobes) have shown the potential to provide a rapid and sensitive detection method for Mtb and single base alterations associated with antibiotic resistance, namely in rpoB gene associated to RIF resistance. We developed a strategy based on the isothermal amplification of sample DNA (LAMP) coupled to specific Au-nanoprobes capable of identifying members of the Mtb complex (MTBC) and discriminating specific mutations within the rpoB gene. Integration of LAMP and Au-nanoprobe assay allowed to detect MTBC member and identify mutations linked to RIF resistance. A total of 12 biological samples were tested and a 100% specificity and sensitivity was attained. There is an increasing demand for simple, fast and cheap methods for the molecular identification of Mtb and for the detection of molecular tags associated to drug resistance suitable for use at point-of-need. Here we describe such a method, that as the potential to get molecular diagnostic of tuberculosis to remote environments.
登录
查看更多内容
影响因子：
2.3
作者：
Doria, G.;Franco, R.;Baptista, P.
通讯作者：
Baptista, P.
影响因子：
168.9
作者：
TELENTI, A;IMBODEN, P;BODMER, T
通讯作者：
BODMER, T
环介导等温扩增后基于金纳米颗粒的目标 DNA 比色检测
影响因子：
3.1
作者：
Zhou Chao;Mu Ying;Jin Qin-han
通讯作者：
Jin Qin-han
影响因子：
9.4
作者：
Iwamoto, T;Sonobe, T;Hayashi, K
通讯作者：
Hayashi, K
影响因子：
3.5
作者：
Veigas, Bruno;Machado, Diana;Baptista, Pedro V.
通讯作者：
Baptista, Pedro V.