Objective: To evaluate the performance of locked nucleic acid (LNA) probe Real-time polymerase chain reaction (PCR) in the detection of Aspergillus fumigatus (A. fumigatus).
Methods: 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.
Results: 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.
Conclusion: LNA probe Real-time PCR is a promisingly accurate assay of rapidly detecting A. fumigatus practically and cost-efficiently.