CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal.
Nanotechnology. 2010 Oct 15;21(41):415101. doi: 10.1088/0957-4484/21/41/415101. Epub 2010 Sep 16.
Tuberculosis (TB) is one of the leading causes of infection in humans, causing high morbility and mortality all over the world. The rate of new cases of multidrug resistant tuberculosis (MDRTB) continues to increase, and since these infections are very difficult to manage, they constitute a serious health problem. In most cases, drug resistance in Mycobacterium tuberculosis has been related to mutations in several loci within the pathogen's genome. The development of fast, cheap and simple screening methodologies would be of paramount relevance for the early detection of these mutations, essential for the timely and effective diagnosis and management of MDRTB patients. The use of gold nanoparticles derivatized with thiol-modified oligonucleotides (Au-nanoprobes) has led to new approaches in molecular diagnostics. Based on the differential non-cross-linking aggregation of Au-nanoprobes, we were able to develop a colorimetric method for the detection of specific sequences and to apply this approach to pathogen identification and single base mutations/single nucleotide polymorphisms (SNP) discrimination. Here we report on the development of Au-nanoprobes for the specific identification of SNPs within the beta subunit of the RNA polymerase (rpoB locus), responsible for resistance to rifampicin in over 95% of rifampicin resistant M. tuberculosis strains.
结核病（TB）是人类感染的主要原因之一，在全球范围内导致高发病率和死亡率。耐多药结核病（MDRTB）的新发病例率持续上升，由于这些感染非常难以治疗，因此构成了严重的健康问题。在大多数情况下，结核分枝杆菌的耐药性与病原体基因组内的几个基因座中的突变有关。开发快速、廉价且简单的筛选方法对于早期检测这些突变至关重要，这对于及时有效地诊断和管理 MDRTB 患者至关重要。使用巯基修饰的寡核苷酸衍生的金纳米粒子（Au-纳米探针）已经为分子诊断学带来了新的方法。基于 Au-纳米探针的差分非交联聚集，我们能够开发出一种用于检测特定序列的比色法，并将这种方法应用于病原体鉴定和单碱基突变/单核苷酸多态性（SNP）的区分。在这里，我们报告了用于特异性鉴定 RNA 聚合酶β亚基（rpoB 基因座）内 SNP 的 Au-纳米探针的开发，该基因座负责超过 95%的利福平耐药结核分枝杆菌菌株对利福平的耐药性。
