Multienzyme isothermal rapid amplification combined with CRISPR/Cas12a for sensitive prostate cancer screening using urine collected without digital rectal examination

IF 4.9 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchemical Journal Pub Date : 2025-03-06 DOI:10.1016/j.microc.2025.113218

Hong Chen , Jiakai Liu , Yongyi Chen , Zhen Song , Jichun Shao , Bo Shi , Yang He

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Abstract

Prostate cancer associated 3 (PCA3), a specific nucleic acid biomarker of prostate cancer, may appear earlier than the prostate-specific antigen (PSA) in prostate cancer. At present, urine, to be used for PCA3 detection, is collected after digital rectal examination (DRE). However, omitting DRE without negatively affecting assay sensitivity may help standardize urine collection procedures and increase PCA3 diagnostic applicability. Based on the DNA replication function of multienzyme isothermal rapid amplification (MIRA) and the specific DNA-shearing function of CRISPR/Cas12a, the MIRA-CRISPR/Cas12a “one-pot” system was developed. The system was shown to detect 0.01 ng/µL RNA in 40 min at room temperature. In addition, this “one-pot” ambient temperature reaction system reduced aerosol contamination and reliance on expensive equipment. Of note, the assay results could be obtained with the naked eye under ultraviolet light. Finally, PCA3 detection results revealed that the marker had great potential for prostate cancer screening, with a specificity of 83.3 %. Compared with the 33.3 % specificity of serum PSA, the specificity of PCA3 for prostate cancer screening was increased by approximately 50 %.

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来源期刊

Microchemical Journal 化学-分析化学

CiteScore

8.70

自引率

8.30%

发文量

1131

审稿时长

1.9 months

期刊介绍： The Microchemical Journal is a peer reviewed journal devoted to all aspects and phases of analytical chemistry and chemical analysis. The Microchemical Journal publishes articles which are at the forefront of modern analytical chemistry and cover innovations in the techniques to the finest possible limits. This includes fundamental aspects, instrumentation, new developments, innovative and novel methods and applications including environmental and clinical field. Traditional classical analytical methods such as spectrophotometry and titrimetry as well as established instrumentation methods such as flame and graphite furnace atomic absorption spectrometry, gas chromatography, and modified glassy or carbon electrode electrochemical methods will be considered, provided they show significant improvements and novelty compared to the established methods.