摘要
目的建立系统性红斑狼疮(SLE)合并白念珠菌和黄曲霉感染的多重荧光定量PCR检测技术,为其诊断提供快速、敏感、特异的方法。方法通过ATCC公布的白念珠菌及黄曲霉的基因序列,应用分子生物学软件设计两对引物和Taqman探针,通过检测狼疮患者合并白念珠菌及黄曲霉等深部真菌感染,怀疑深部真菌感染样品各20例,以及非白念珠菌及非黄曲霉微生物样品20例,以评价该法阳性率、敏感性、特异性。结果多重荧光定量PCR检测SLE合并白念珠菌和黄曲霉感染的阳性率及特异性均为100%;而该法和真菌培养法敏感性分别为75.00%和40.00%,且有统计学差异(P<0.05)。结论基于Taqman探针技术的实时荧光PCR检测体系,可同时检测SLE患者合并白念珠菌和黄曲霉深部真菌感染,具有快速、灵敏度高、特异性强、可定量等优点。
Objective To establish a rapid, sensitive and specific method based on multiplex fluorescent quantitative PCR for detection of deep infections with Candida albicans and A spergillus flavus in patients with systemic lupus erythematosus （SLE）. Methods Two pairs of primers and Taqman probes were designed according to the gene sequences of Condida albicons and Aspergillus flavus available in American Type Culture Collection. The positivity rate, sensitivity and specificity of the multiplex fluorescent quantitative PCR-based method for detecting the fungal infection was tested in 20 specimens from SLE patients with Candida albicans and AspergiUus flavus infections, 20 specimens from SLE patients with suspected deep fungal infections, and 20 microbial samples other than Condida albicons or Aspergillus flavus. Results The multiple fluorescence quantitative PCR-based method showed a positivity rate and specificity of both 100% for detecting Candida albicans and AspergillusflavusinfecfionsintheSLEpatients. This method resulted in a detection sensitivity of 75%, significantly higher than that of fugal culture method （40%, P〈0.05）. Conclusions The multiplex fluorescent real-time PCR-based method allows rapid, quantitative and simultaneous detection of deep Candida albicans and Aspergillus flavus infections with high sensitivity and specificity in SLE patients.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2009年第10期2112-2114,2121,共4页
Journal of Southern Medical University
关键词
系统性红斑狼疮
深部真菌
聚合酶链反应
systemic lupus erythematosus
deep fungal infection
polymerase chain reaction
