Introduction: (, CA) is an essential invasive fungus in clinical diagnosis. Although several detection methods exist, none meet the need for early diagnosis. A rapid, sensitive, and specific diagnostic tool is crucial for effective prevention and control of infections.
Methods: This study aimed to develop a new, rapid, and ultrasensitive diagnostic tool for detection based on restriction endonuclease-mediated real-time loop-mediated isothermal amplification (ERT-LAMP-CA). The ERT-LAMP-CA technology combines LAMP amplification, restriction endonuclease cleavage, and real-time fluorescence detection in a single reaction tube, which can complete a diagnosis of in a short time (approximately 1 h).
Results: Herein, we developed the primer sequences required for ERT-LAMP-CA based on the ITS2 gene of and found that ERT-LAMP-CA limit of detection was approximately 500 ag/μL genomic DNA and can present negative results for non- templates. We tested sputum samples from 64 patients with suspected infections to validate ERT-LAMP-CA applicability in clinical sample testing and found that ERT-LAMP-CA was consistent with multiplex PCR-capillary electrophoresis.
Discussion: In conclusion, ERT-LAMP-CA is a rapid, accurate, and sensitive assay with excellent potential for clinical and basic laboratory diagnosis and an efficient screening strategy.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11586279 | PMC |
| http://dx.doi.org/10.3389/fcimb.2024.1450199 | DOI Listing |
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