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To promptly identify common respiratory viruses causing infections and facilitate effective therapeutic interventions in resource-limited settings such as grassroots, clinics, border areas, and border defense, this study established a visual detection method based on recombinase polymerase amplification (RPA) technology and CRISPR/Cas12a (clustered regularly interspaced short palindromic repeats/CRISPR-associated 12a) system.
Initially, conservative sequences of each virus target were designed, and RPA primers along with single-strand guide RNAs (sgRNAs) were selected. Subsequently, the RPA technology was integrated with the CRISPR/Cas12a detection method for visual detection of influenza A virus, influenza B virus, respiratory syncytial virus, Severe Acute Respiratory Syndrome Coronavirus 2, and human rhinovirus.
The detection method yielded results within 1.5 hours, with a sensitivity of 3.5 copies/μl plasmid, and exhibited no cross-reactivity between each virus target. In terms of detection accuracy, this method demonstrated higher consistency compared to the control Quantitative Reverse Transcription Polymerase Chain Reaction, (qRT-PCR) method.
The visual detection method established in this study possesses good specificity and high sensitivity for the common five respiratory virus infections. It is suitable for on-site detection of common respiratory virus infections, especially in resource-limited environments, and holds promising clinical application prospects.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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