Detection of medically important Candida species by absolute quantitation real-time polymerase chain reaction

Jundishapur Journal of Microbiology
Leslie Thian Lung ThanHeng Fong Seow

Abstract

The number of invasive candidiasis cases has risen especially with an increase in the number of immunosuppressed and immunocom promised patients. The early detection of Candida species which is specific and sensitive is important in determining the correct administration of antifungal drugs to patients. This study aims to develop a method for the detection, identification and quantitation of medically important Candida species through quantitative polymerase chain reaction (qPCR). The isocitrate lyase (ICL) gene which is not found in mammals was chosen as the target gene of real-time PCR. Absolute quantitation of the gene copy number was achieved by constructing the plasmid containing the ICL gene which is used to generate standard curve. Twenty fungal species, two bacterial species and human DNA were tested to check the specificity of the detection method. All eight Candida species were successfully detected, identified and quantitated based on the ICL gene. A seven-log range of the gene copy number and a minimum detection limit of 10(3) copies were achieved. A one-tube absolute quantification real-time PCR that differentiates medically important Candida species via individual unique melting temperature was achieved. Analytica...Continue Reading

References

Jul 14, 2001·Nature·M C Lorenz, G R Fink
Jun 18, 2002·Trends in Molecular Medicine·Dieter Klein
Feb 17, 2006·Molecular Aspects of Medicine·F WatzingerT Lion
Nov 27, 2007·Diagnostic Microbiology and Infectious Disease·Catherine DunyachMichele Mallié
Dec 14, 2011·Diagnostic Microbiology and Infectious Disease·Leslie Thian Lung ThanHeng Fong Seow
May 15, 2012·Diagnostic Microbiology and Infectious Disease·Daniel DiekemaMichael Pfaller

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

CLUSTALW
BLAST

Related Concepts

Related Feeds

Antifungals

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.

Antifungals (ASM)

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.

Candida albicans

Candida albicans is an opportunistic, fungal pathogen of humans that frequently causes superficial infections of oral and vaginal mucosal surfaces of debilitated and susceptible individuals. Discover the latest research on Candida albicans here.