| 白色念珠菌ITS2的实时荧光定量PCR快速检测方法的建立及评价 |
| 投稿时间：2015-08-05 修订日期：2015-09-14 点此下载全文 |
| 引用本文：郭毅,杨靖娴,邵冬华,刘静,梁国威.白色念珠菌ITS2的实时荧光定量PCR快速检测方法的建立及评价[J].医学研究杂志,2016,45(7):79-84 |
| DOI：
10.11969/j.issn.1673-548X.2016.07.022 |
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| 基金项目:北京大学航天临床医学院科研基金资助项目（YN201322） |
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| 中文摘要:目的 建立一种快速鉴定白色念珠菌的实时荧光定量PCR检测方法。方法 在白色念珠菌核糖体RNA编码基因（rDNA）中的内转录间隔区2（ITS2）上设计特异性引物和TaqMan-MGB探针，通过实时荧光定量PCR分析系统建立白色念珠菌的检测方法，对其敏感度、重复性和特异性进行方法学评价，并在模拟白色念珠菌血症的血标本中初步探讨其应用价值。结果 所建方法对白色念珠菌纯菌液的准确检测下限为101CFU/ml，相应的批内、批间变异系数（CV）分别为1.57%和2.20%。对135株白色念珠菌临床分离株、其他非白念菌株、细菌临床分离株和血液标本进行检测，结果显示该方法特异性为100%。另外，对于模拟白色念珠菌血症的血标本，该方法的最低检测下限为102CFU/ml。结论 所建方法具有特异性和敏感度高、重复性好的特性，可适用于临床上白色念珠菌引起的血流感染的快速鉴定。 |
| 中文关键词:白色念珠菌|实时荧光定量PCR|内转录间隔区2(ITS 2)|血流感染 |
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| A Novel Real-time PCR Assay Based on the Internal Transcribed Spacer 2 (ITS2) for Rapid Identification of Candida Albicans |
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| Abstract:Objective To develop a novel real-time PCR for rapid identification of Candida albicans(C. albicans). Methods Unique primers and the TaqMan-MGB probe were designed on the internal transcribed spacer 2 (ITS2)of rRNA gene(rDNA) in C. albicans. The sensitivity, specificity and reproducibility of the method were then evaluated. Meanwhile, clinical applicability to detect candidemia of the assay was tested in simulated C. albicans infection blood samples. Results The lowest amount of C. albicans that the assay can measure accurately in suspensions was 101CFU/ml. The coefficients of variability(CV) for the detection limit were 1.57% and 2.20% in inter-assay and intra-assay respectively. The reaction specificity was 100% when evaluating the assay using DNA samples from clinical isolates of 135 C. albicans strains, other Candida spp. strains, bacterial strains and human blood. For simulated infection blood samples, the detection limit of C. albicans was 102 CFU/ml. Conclusion The newly developed method was a highly sensitive and specific assay, which could also be applied to rapid diagnosis of blood stream infection induced by C. albicans. |
| keywords:Candida albicans|Quantitative real-time PCR|Internal transcribed spacer 2(ITS2)|Blood stream infection(BSI) |
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