A Rapid, Highly Sensitive and Open-Access SARS-CoV-2 Detection Assay for Laboratory and Home Testing

被引:24
作者
Kellner, Max J. [1 ,2 ,3 ,4 ]
Ross, James J. [2 ,3 ]
Schnabl, Jakob [2 ,3 ]
Dekens, Marcus P. S. [1 ]
Matl, Martin [1 ,2 ]
Heinen, Robert [1 ,2 ]
Grishkovskaya, Irina [1 ]
Bauer, Benedikt [1 ]
Stadlmann, Johannes [1 ,2 ,5 ]
Menendez-Arias, Luis [6 ]
Straw, Andrew D. [7 ,8 ]
Fritsche-Polanz, Robert [9 ]
Traugott, Marianna
Seitz, Tamara
Zoufaly, Alexander
Foedinger, Manuela [9 ]
Wenisch, Christoph
Zuber, Johannes [1 ,10 ,11 ]
Pauli, Andrea [1 ]
Brennecke, Julius [2 ]
机构
[1] Vienna Bioctr VBC, Res Inst Mol Pathol IMP, Vienna, Austria
[2] Inst Mol Biotechnol Austrian Acad Sci IMBA, Vienna Bioctr VBC, Vienna, Austria
[3] Doctoral Sch Univ Vienna, Med Univ Vienna, Vienna Bioctr PhD Program, Vienna, Austria
[4] MRC Lab Mol Biol, Cambridge, England
[5] Univ Nat Resources & Life Sci, Dept Chem, Vienna, Austria
[6] Univ Autonoma Madrid, Consejo Super Invest Cient, Ctr Biol Mol Severo Ochoa, Madrid, Spain
[7] Albert Ludwigs Univ Freiburg, Inst Biol 1, Fac Biol, Freiburg, Germany
[8] Albert Ludwigs Univ Freiburg, Bernstein Ctr Freiburg, Fac Biol, Freiburg, Germany
[9] Inst Lab Diagnost, Vienna, Austria
[10] 4th Med Dept Infect Dis & Trop Med, Vienna, Austria
[11] Med Univ Vienna, Vienna Bioctr VBC, Vienna, Austria
基金
奥地利科学基金会; 欧洲研究理事会;
关键词
COVID-19; diagnostics; RT-LAMP; coronavirus; SARS-CoV-2; isothermal amplification; open-source; MEDIATED ISOTHERMAL AMPLIFICATION; REVERSE-TRANSCRIPTASE; DNA-POLYMERASE; PURIFICATION; FIDELITY; LAMP;
D O I
10.3389/fmolb.2022.801309
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
RT-qPCR-based diagnostic tests play important roles in combating virus-caused pandemics such as Covid-19. However, their dependence on sophisticated equipment and the associated costs often limits their widespread use. Loop-mediated isothermal amplification after reverse transcription (RT-LAMP) is an alternative nucleic acid detection method that overcomes these limitations. Here, we present a rapid, robust, and sensitive RT-LAMP-based SARS-CoV-2 detection assay. Our 40-min procedure bypasses the RNA isolation step, is insensitive to carryover contamination, and uses a colorimetric readout that enables robust SARS-CoV-2 detection from various sample types. Based on this assay, we have increased sensitivity and scalability by adding a nucleic acid enrichment step (Bead-LAMP), developed a version for home testing (HomeDip-LAMP), and identified open-source RT-LAMP enzymes that can be produced in any molecular biology laboratory. On a dedicated website, (DOI: ), we provide detailed protocols and videos. Our optimized, general-purpose RT-LAMP assay is an important step toward population-scale SARS-CoV-2 testing.
引用
收藏
页数:21
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