Julia Alcoba-Florez1, Rafaela González-Montelongo2, Antonio Íñigo-Campos3, Diego García-Martínez de Artola4, Helena Gil-Campesino5, Laura Ciuffreda6, Agustín Valenzuela-Fernández7, Carlos Flores8. 1. Servicio de Microbiología, Hospital Universitario N. S. de Candelaria, 38010 Santa Cruz de Tenerife, Spain. Electronic address: juliaalcoba@gmail.com. 2. Genomics Division, Instituto Tecnológico y de Energías Renovables, 38600 Santa Cruz de Tenerife, Spain. Electronic address: rgonzalezmontelongo@iter.es. 3. Genomics Division, Instituto Tecnológico y de Energías Renovables, 38600 Santa Cruz de Tenerife, Spain. Electronic address: ainigo@iter.es. 4. Servicio de Microbiología, Hospital Universitario N. S. de Candelaria, 38010 Santa Cruz de Tenerife, Spain. Electronic address: diegogarciamartinezdeartola@gmail.com. 5. Servicio de Microbiología, Hospital Universitario N. S. de Candelaria, 38010 Santa Cruz de Tenerife, Spain. Electronic address: helegc@hotmail.com. 6. Research Unit, Hospital Universitario N. S. de Candelaria, 38010 Santa Cruz de Tenerife, Spain. Electronic address: laura.ciuffreda1988@gmail.com. 7. Laboratorio de Inmunología Celular y Viral, Unidad de Farmacología, Facultad de Medicina, Universidad de La Laguna, 38200 San Cristóbal de La Laguna, Spain; Red española de Investigación en VIH/SIDA (RIS)-RETIC, Instituto de Salud Carlos III, 28029 Madrid, Spain. Electronic address: avalenzu@ull.edu.es. 8. Genomics Division, Instituto Tecnológico y de Energías Renovables, 38600 Santa Cruz de Tenerife, Spain; Research Unit, Hospital Universitario N. S. de Candelaria, 38010 Santa Cruz de Tenerife, Spain; CIBER de Enfermedades Respiratorias, Instituto de Salud Carlos III, 28029 Madrid, Spain; Instituto de Tecnologías Biomédicas (ITB) Universidad de La Laguna, 38200 San Cristóbal de La Laguna, Spain. Electronic address: cflores@ull.edu.es.
Abstract
OBJECTIVES: The gold-standard COVID-19 diagnosis relies on detecting SARS-CoV-2 using RNA purification and one-step retrotranscription and quantitative PCR (RT-qPCR). Based on the urgent need for high-throughput screening, we tested the performance of three alternative, simple and affordable protocols to rapidly detect SARS-CoV-2, bypassing the long and tedious RNA extraction step and reducing the time to viral detection. METHODS: We evaluated three methods based on direct nasopharyngeal swab viral transmission medium (VTM) heating before the RT-qPCR: a) direct without additives; b) in a formamide-EDTA (FAE) buffer, c) in a RNAsnapTM buffer. RESULTS: Although with a delay in cycle threshold compared to the gold-standard, we found consistent results in nasopharyngeal swab samples that were subject to a direct 70°C incubation for 10 min. CONCLUSIONS: Our findings provide valuable options to overcome any supply chain issue and help to increase the throughput of diagnostic tests, thereby complementing standard diagnosis.
OBJECTIVES: The gold-standard COVID-19 diagnosis relies on detecting SARS-CoV-2 using RNA purification and one-step retrotranscription and quantitative PCR (RT-qPCR). Based on the urgent need for high-throughput screening, we tested the performance of three alternative, simple and affordable protocols to rapidly detect SARS-CoV-2, bypassing the long and tedious RNA extraction step and reducing the time to viral detection. METHODS: We evaluated three methods based on direct nasopharyngeal swab viral transmission medium (VTM) heating before the RT-qPCR: a) direct without additives; b) in a formamide-EDTA (FAE) buffer, c) in a RNAsnapTM buffer. RESULTS: Although with a delay in cycle threshold compared to the gold-standard, we found consistent results in nasopharyngeal swab samples that were subject to a direct 70°C incubation for 10 min. CONCLUSIONS: Our findings provide valuable options to overcome any supply chain issue and help to increase the throughput of diagnostic tests, thereby complementing standard diagnosis.
Authors: Julia Alcoba-Florez; Helena Gil-Campesino; Diego García-Martínez de Artola; Oscar Díez-Gil; Agustín Valenzuela-Fernández; Rafaela González-Montelongo; Laura Ciuffreda; Carlos Flores Journal: Int J Infect Dis Date: 2020-11-18 Impact factor: 3.623
Authors: José P Miranda; Javiera Osorio; Mauricio Videla; Gladys Angel; Rossana Camponovo; Marcela Henríquez-Henríquez Journal: Front Med (Lausanne) Date: 2020-10-15
Authors: Emily A Bruce; Meei-Li Huang; Garrett A Perchetti; Scott Tighe; Pheobe Laaguiby; Jessica J Hoffman; Diana L Gerrard; Arun K Nalla; Yulun Wei; Alexander L Greninger; Sean A Diehl; David J Shirley; Debra G B Leonard; Christopher D Huston; Beth D Kirkpatrick; Julie A Dragon; Jessica W Crothers; Keith R Jerome; Jason W Botten Journal: PLoS Biol Date: 2020-10-02 Impact factor: 8.029