Real‐Time PCR Assay to Detect DNA in Sera for the Diagnosis of Deep‐Seated Trichosporonosis

Mekha, Nanthawan; Sugita, Takashi; Ikeda, Reiko; Nishikawa, Akemi; Poonwan, Natteewan

doi:10.1111/j.1348-0421.2007.tb03950.x

Cited by 4 publications

(1 citation statement)

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“…There have also been reports on the design of species-specific primers for real-time quantitative PCR based on the IGS1 region for the detection of T. asahii in whole blood samples. However, the procedure required extraction of DNA from blood samples for amplification, which increased the total time of detection [29]. Our previous study on the detection of T. asahii using LAMP technology also showed requirement for DNA extraction after obtaining pure culture, and the cost of this technology was much higher than that of PCR, both of which hindered the application of this technology in clinical practice [8].…”

Section: Discussionmentioning

confidence: 99%

Rapid and Simple Detection ofTrichosporon asahiiby Optimized Colony PCR

Zhang

Liao³

et al. 2019

BioMed Research International

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Trichosporon asahii is the major pathogen causing invasive trichosporonosis. Conventional methods of its detection are time-consuming or costly and often require complex DNA extraction and purification steps, which hinders rapid clinical diagnosis. In this study, we evaluated colony PCR, which directly uses colonies or trace clinical samples as the template for amplification, for rapid detection of T. asahii infection. Four methods, namely, direct colony, freeze-thaw, glass beads, and enzymolysis, were compared to select the best DNA extraction strategy. We subsequently designed and screened species-specific primers targeting the intergenic spacer 1 (IGS1) of the ribosomal DNA of T. asahii and used them to detect mock infection clinical samples. The species-specific colony PCR based on glass beads proved advantageous, with short procedure time (154.8 ± 0.6 min), good sensitivity (detection limit, 102 CFU/mL), and specificity for T. asahii, indicating that this method can be used for the rapid and simple identification of clinical samples of T. asahii infection.

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Section: Discussionmentioning

confidence: 99%