{"doi":"10.1371/journal.pone.0245164","title":"Development of a reverse transcription recombinase polymerase amplification assay for rapid and direct visual detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)","abstract":"Rapid diagnosis is an important intervention in managing the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) outbreak. Real time reverse transcription polymerase chain reaction (RT-qPCR) remains the primary means for diagnosing the new virus strain but it is time consuming and costly. Recombinase polymerase amplification (RPA) is an isothermal amplification assay that does not require a PCR machine. It is an affordable, rapid, and simple assay. In this study, we developed and optimized a sensitive reverse transcription (RT)-RPA assay for the rapid detection of SARS-CoV-2 using SYBR Green I and/or lateral flow (LF) strip. The analytical sensitivity and specificity of the RT-RPA assay were tested by using 10-fold serial diluted synthetic RNA and genomic RNA of similar viruses, respectively. Clinical sensitivity and specificity of the RT-RPA assay were carried out using 78 positive and 35 negative nasopharyngeal samples. The detection limit of both RPA and RT-qPCR assays was 7.659 and 5 copies/μL RNA, respectively with no cross reactivity with other viruses. The clinical sensitivity and specificity of RT-RPA were 98% and 100%, respectively. Our study showed that RT-RPA represents a viable alternative to RT-qPCR for the detection of SARS-CoV-2, especially in areas with limited infrastructure.","journal":"PLoS ONE","year":2021,"id":154978,"datarank":0.62147020895873,"base_score":4.143134726391533,"endowment":4.143134726391533,"self_citation_contribution":0.62147020895873,"citation_network_contribution":0.0,"self_endowment_contribution":0.62147020895873,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":62,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.958,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":657622,"name":"Ilyiana Ismail","orcid":null,"position":1,"is_corresponding":false},{"id":657623,"name":"Nur Izati Mustapa","orcid":null,"position":2,"is_corresponding":false},{"id":656551,"name":"Meng Yee Lai","orcid":"0000-0002-4544-695X","position":3,"is_corresponding":false},{"id":657624,"name":"Tuan Suhaila Tuan Soh","orcid":null,"position":4,"is_corresponding":false},{"id":657625,"name":"Afifah Haji Hassan","orcid":null,"position":5,"is_corresponding":false},{"id":266015,"name":"Kalaiarasu M. Peariasamy","orcid":"0000-0001-9279-3498","position":6,"is_corresponding":false},{"id":656552,"name":"Yee Leng Lee","orcid":"0000-0001-6944-3836","position":7,"is_corresponding":false},{"id":656553,"name":"Maria Kahar Bador","orcid":"0000-0002-0881-133X","position":8,"is_corresponding":false},{"id":657626,"name":"Jennifer Chong","orcid":null,"position":9,"is_corresponding":false},{"id":269033,"name":"Pik Pin Goh","orcid":null,"position":10,"is_corresponding":false},{"id":656550,"name":"Yee Ling Lau","orcid":"0000-0002-2037-2165","position":0,"is_corresponding":true}],"reference_count":15,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T23:43:59.154159Z","pmid":"33406112","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":[]}