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10.3389/fcimb.2020.00331

http://scihub22266oqcxt.onion/10.3389/fcimb.2020.00331
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suck abstract from ncbi


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pmid32626666      Front+Cell+Infect+Microbiol 2020 ; 10 (ä): 331
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  • Development and Validation of a Rapid, Single-Step Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP) System Potentially to Be Used for Reliable and High-Throughput Screening of COVID-19 #MMPMID32626666
  • Jiang M; Pan W; Arasthfer A; Fang W; Ling L; Fang H; Daneshnia F; Yu J; Liao W; Pei H; Li X; Lass-Florl C
  • Front Cell Infect Microbiol 2020[]; 10 (ä): 331 PMID32626666show ga
  • Objectives: Development and validation of a single-step and accurate reverse transcriptase loop-mediated isothermal amplification technique (RT-LAMP) for rapid identification of SARS-CoV-2 relative to commercial quantitative reverse transcriptase real-time PCR (qRT-PCR) assays to allow prompt initiation of proper medical care and containment of virus spread. Methods: Primers showing optimal in-silico features were subjected to analytical sensitivity and specificity to assess the limit of detection (LOD) and cross-reaction with closely- and distantly-related viral species, and clinically prominent bacterial and fungal species. In order to evaluate the clinical utility, our RT-LAMP was subjected to a large number of clinical samples, including 213 negative and 47 positive patients, relative to two commercial quantitative RT-PCR assays. Results: The analytical specificity and sensitivity of our assay was 100% and 500 copies/ml when serial dilution was performed in both water and sputum. Subjecting our RT-LAMP assay to clinical samples showed a high degree of specificity (99.5%), sensitivity (91.4%), positive predictive value (97.7%), and negative predictive value (98.1%) when used relative to qRT-PCR. Our RT-LAMP assay was two times faster than qRT-PCR and is storable at room temperature. A suspected case that later became positive tested positive using both our RT-LAMP and the two qRT-PCR assays, which shows the capability of our assay for screening purposes. Conclusions: We present a rapid RT-LAMP assay that could extend the capacity of laboratories to process two times more clinical samples relative to qRT-PCR and potentially could be used for high-throughput screening purposes when demand is increasing at critical situations.
  • |*Nucleic Acid Amplification Techniques[MESH]
  • |Betacoronavirus/*genetics[MESH]
  • |COVID-19[MESH]
  • |Coronavirus Infections/*diagnosis[MESH]
  • |Humans[MESH]
  • |Mass Screening/*methods[MESH]
  • |Pandemics[MESH]
  • |Pneumonia, Viral/*diagnosis[MESH]
  • |Real-Time Polymerase Chain Reaction[MESH]
  • |Reverse Transcriptase Polymerase Chain Reaction[MESH]
  • |SARS-CoV-2[MESH]


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