Development of a Luminex-Based Multiplex Assay for Detection of Mutations Conferring Resistance to Echinocandins in Candida glabrata
Development of a Luminex-Based Multiplex Assay for Detection of Mutations Conferring Resistance to Echinocandins in Candida glabrata
复制标题
DOI：
10.1128/jcm.03378-13
复制
发表时间：
2014-03-01
影响因子：
9.4
通讯作者：
Lockhart, Shawn R.
中科院分区：
文献类型：
--
作者：
Pham, Cau D.;Bolden, Carol B.;Lockhart, Shawn R.
棘白菌素是光滑念珠菌引起的侵袭性念珠菌病的推荐治疗方法。已知对棘白菌素的耐药性是由FKS 1和FKS 2基因的热点-1（HS 1）区域的非同义突变引起的，FKS 1和FKS 2基因编码棘白菌素的靶标β-1，3-葡聚糖合酶的亚基。在这里，我们描述了一种基于微球的检测方法的开发，该方法使用Luminex MagPix技术来鉴定FKS 1 HS 1和FKS 2 HS 1结构域中的突变，这些突变在体外赋予C.光果苔属分离物该测定是快速的，并且可以以高通量进行。使用先前通过DNA测序表征的具有FKS 1 HS 1和FKS 2 HS 1结构域的102株分离株对该测定法进行了验证。该测定与DNA测序结果100%一致。然后将该测定用于1，032 C的高通量筛选。光滑果属监测分离株。鉴定了16个新的突变分离株，包括我们收集的新突变（del 659 F）。该检测方法提供了一种快速、经济的方法来筛选C。光滑分离物的棘白菌素抗性。
Echinocandins are the recommended treatment for invasive candidiasis due to Candida glabrata. Resistance to echinocandins is known to be caused by nonsynonymous mutations in the hot spot-1 (HS1) regions of the FKS1 and FKS2 genes, which encode a subunit of the beta-1,3-glucan synthase, the target of echinocandins. Here, we describe the development of a microsphere-based assay using Luminex MagPix technology to identify mutations in the FKS1 HS1 and FKS2 HS1 domains, which confer in vitro echinocandin resistance in C. glabrata isolates. The assay is rapid and can be performed with high throughput. The assay was validated using 102 isolates that had FKS1 HS1 and FKS2 HS1 domains previously characterized by DNA sequencing. The assay was 100% concordant with DNA sequencing results. The assay was then used for high-throughput screening of 1,032 C. glabrata surveillance isolates. Sixteen new isolates with mutations, including a mutation that was new to our collection (del659F), were identified. This assay provides a rapid and cost-effective way to screen C. glabrata isolates for echinocandin resistance.