Ontology highlight
ABSTRACT: Background
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.Objectives
This study aims to develop a method for the detection, identification and quantitation of medically important Candida species through quantitative polymerase chain reaction (qPCR).Materials and methods
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.Results
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.Conclusions
A one-tube absolute quantification real-time PCR that differentiates medically important Candida species via individual unique melting temperature was achieved. Analytical sensitivity and specificity were not compromised.
SUBMITTER: Than LT
PROVIDER: S-EPMC4350049 | biostudies-literature | 2015 Jan
REPOSITORIES: biostudies-literature
Than Leslie Thian Lung LT Chong Pei Pei PP Ng Kee Peng KP Seow Heng Fong HF
Jundishapur journal of microbiology 20141210 1
<h4>Background</h4>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.<h4>Objectives</h4>This study aims to develop a method for the detection, identification and quantitation of medically important Candida species through quantitative polymerase ...[more]