[Multiplex fluorescent quantitative PCR for detecting deep fungal infection in patients with systemic lupus erythematosus].
OBJECTIVE
To establish a rapid, sensitive and specific method based on multiplex fluorescent quantitative PCR for detection of deep infections with Candida albicans and Aspergillus flavus in patients with systemic lupus erythematosus (SLE).


METHODS
Two pairs of primers and Taqman probes were designed according to the gene sequences of Candida albicans 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 Aspergillus flavus infections, 20 specimens from SLE patients with suspected deep fungal infections, and 20 microbial samples other than Candida albicans 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 Aspergillus flavus infections in the SLE patients. 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.
Chen
Ming-yu
MY
Department of Dermatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. chenmingyu3w@163.com
Sun
Le-dong
LD
Zhao
Jia
J
Zeng
Kang
K
D000328:Adult; D001228:Aspergillosis / Q000150:complications / Q000175:diagnosis / Q000382:microbiology; D001230:Aspergillus / Q000235:genetics / Q000302:isolation & purification*; D002176:Candida albicans / Q000235:genetics / Q000302:isolation & purification*; D002177:Candidiasis / Q000150:complications / Q000175:diagnosis / Q000382:microbiology; D002681:China / Q000453:epidemiology; D005260:Female; D005453:Fluorescence; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000150:complications / Q000382:microbiology*; D008297:Male; D009181:Mycoses / Q000150:complications / Q000175:diagnosis* / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity