{"doi":"10.1016/j.bios.2023.115866","title":"Sensitive and specific CRISPR-Cas12a assisted nanopore with RPA for Monkeypox detection","abstract":"Monkeypox virus (MPXV) poses a global health emergency, necessitating rapid, simple, and accurate detection to manage its spread effectively. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technique has emerged as a promising next-generation molecular diagnostic approach. Here, we developed a highly sensitive and specific CRISPR-Cas12a assisted nanopore (SCAN) with isothermal recombinase polymerase amplification (RPA) for MPXV detection. The RPA-SCAN method offers a sensitivity unachievable with unamplified SCAN while also addressing the obstacles of PCR-SCAN for point-of-care applications. We demonstrated that size-counting of single molecules enables analysis of reaction-time dependent distribution of the cleaved reporter. Our MPXV-specific RPA assay achieved a limit of detection (LoD) of 19 copies in a 50 μL reaction system. By integrating 2 μL of RPA amplifications into a 20 μL CRISPR reaction, we attained an overall LoD of 16 copies/μL (26.56 aM) of MPXV at a 95% confidence level using the SCAN sensor. We also verified the specificity of RPA-SCAN in distinguishing MPXV from cowpox virus with 100% accuracy. These findings suggest that the isothermal RPA-SCAN device is well-suited for highly sensitive and specific Monkeypox detection. Given its electronic nature and miniaturization potential, the RPA-SCAN system paves the way for diagnosing a wide array of other infectious pathogens at the point of care.","journal":"Biosensors and Bioelectronics","year":2023,"id":318009,"datarank":0.6394019815561974,"base_score":4.2626798770413155,"endowment":4.2626798770413155,"self_citation_contribution":0.6394019815561974,"citation_network_contribution":0.0,"self_endowment_contribution":0.6394019815561974,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":70,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9555,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1024501,"name":"Muhammad Asad Ullah Khalid","orcid":"0000-0001-5926-1764","position":1,"is_corresponding":false},{"id":628287,"name":"Ming Dong","orcid":"0000-0002-9071-5717","position":2,"is_corresponding":false},{"id":886738,"name":"Anthony J. Politza","orcid":"0009-0005-8670-2357","position":3,"is_corresponding":false},{"id":1024502,"name":"Zhikun Zhang","orcid":"0000-0002-0945-7176","position":4,"is_corresponding":false},{"id":628289,"name":"Aneesh Kshirsagar","orcid":"0000-0003-2288-5439","position":5,"is_corresponding":false},{"id":628288,"name":"Tianyi Liu","orcid":"0009-0007-8084-4690","position":6,"is_corresponding":false},{"id":270357,"name":"Weihua Guan","orcid":"0000-0002-8435-9672","position":7,"is_corresponding":false},{"id":1024500,"name":"Md. Ahasan Ahamed","orcid":"0009-0006-9962-2994","position":0,"is_corresponding":true}],"reference_count":31,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T01:06:55.884740Z","pmid":"38029710","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}