Performance of locked nucleic acid probe real-time polymerase chain reaction in the detection of Aspergillus fumigatus

Abstract

To evaluate the performance of locked nucleic acid (LNA) probe Real-time polymerase chain reaction (PCR) in the detection of Aspergillus fumigatus (A. fumigatus). All clinically cultured isolates of Aspergillus at our hospital were identified by morphology and DNA sequencing assay. The experimental group consists of A. fumigatus (n = 48) while the control group was made up of A. flavus (n = 55), A. versicolor (n = 16), A. nidulans (n = 10), A. sydowii (n = 5) and A. parasiticus (n = 1). The clinical samples consisted of A. fumigatus sinusitis tissue (n = 20) and bronchoalveolar lavage fluid (n = 1). DNA was extracted from all samples. A. fumigatus β-tubulin gene was targeted with LNA probe Real-time PCR assay. LNA probe Real-time PCR was evaluated with regards to specificity, efficiency, linear dynamic range in PCR amplification and limits of detection. All clinical samples were positively amplified. The specificity was 100% and the PCR efficiency 98.2%. Linear dynamic range was at least six orders of magnitude and the limit of detection 2.5 pg. LNA probe Real-time PCR is a promisingly accurate assay of rapidly detecting A. fumigatus practically and cost-efficiently.