Monochrome LightCycler PCR assay for detection and quantification of five common species of Candida and Aspergillus.
Invasive fungal pathogens, especially in immunocompromised hosts, can result in life-threatening infections. Current laboratory/radiological methods for fungal identification are time-consuming and lack sensitivity and specificity. A monochrome, multiplex, real-time PCR assay for the identification and quantification of Candida albicans, Candida krusei, Candida tropicalis, Aspergillus flavus and Aspergillus fumigatus is described here. Detection of each of these fungi was specific and demonstrated 100 % concordance with biochemical/culture identification in all 60 isolates tested. Samples from 16 febrile neutropenic patients with haematological malignancies were also analysed and the utility of the assay in clinical samples was reconfirmed without false-negative results. The sensitivity of this assay was 0.1 pg fungal genomic DNA, corresponding to three cells, for C. albicans, C. krusei, C. tropicalis and A. flavus, and 0.01 pg fungal genomic DNA, i.e. less than one cell, for A. fumigatus. The analysis allows a low-cost, simple, rapid and sensitive alternative for clinical identification and quantification of these five common fungal species.
Bu
Rong
R
King Fahad National Center for Children's Cancer and Research, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia 2King Fasial Specialist Hospital and Research Center, Riyadh, Saudi Arabia 3International Network for Cancer Treatment and Research, Brussels, Belgium.
Sathiapalan
Rajeev K
RK
King Fahad National Center for Children's Cancer and Research, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia 2King Fasial Specialist Hospital and Research Center, Riyadh, Saudi Arabia 3International Network for Cancer Treatment and Research, Brussels, Belgium.
Ibrahim
Muna M
MM
King Fahad National Center for Children's Cancer and Research, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia 2King Fasial Specialist Hospital and Research Center, Riyadh, Saudi Arabia 3International Network for Cancer Treatment and Research, Brussels, Belgium.
Al-Mohsen
Ibrahim
I
King Fahad National Center for Children's Cancer and Research, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia 2King Fasial Specialist Hospital and Research Center, Riyadh, Saudi Arabia 3International Network for Cancer Treatment and Research, Brussels, Belgium.
Almodavar
Edna
E
King Fahad National Center for Children's Cancer and Research, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia 2King Fasial Specialist Hospital and Research Center, Riyadh, Saudi Arabia 3International Network for Cancer Treatment and Research, Brussels, Belgium.
Gutierrez
Marina I
MI
King Fahad National Center for Children's Cancer and Research, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia 2King Fasial Specialist Hospital and Research Center, Riyadh, Saudi Arabia 3International Network for Cancer Treatment and Research, Brussels, Belgium.
Bhatia
Kishor
K
King Fahad National Center for Children's Cancer and Research, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia 2King Fasial Specialist Hospital and Research Center, Riyadh, Saudi Arabia 3International Network for Cancer Treatment and Research, Brussels, Belgium.
D001228:Aspergillosis / Q000175:diagnosis / Q000382:microbiology; D001231:Aspergillus flavus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D001232:Aspergillus fumigatus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002175:Candida / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002176:Candida albicans / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D041022:Candida tropicalis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D002177:Candidiasis / Q000175:diagnosis / Q000382:microbiology; D017931:DNA Primers / Q000737:chemistry; D004271:DNA, Fungal / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D004271:DNA, Fungal