An Ultra-Rapid Real-Time RT-PCR Method Using the PCR1100 to Detect Severe Acute Respiratory Syndrome Coronavirus-2
The disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in Wuhan, China, in December 2019, has rapidly spread worldwide. SARS-CoV-2 is usually detected via real-time reverse-transcription polymerase chain reaction (RT-PCR). However, the increase in specimen load in institu...
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| Published in | Japanese Journal of Infectious Diseases Vol. 74; no. 1; pp. 29 - 34 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
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Japan
National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee
29.01.2021
Japan Science and Technology Agency |
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| Abstract | The disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in Wuhan, China, in December 2019, has rapidly spread worldwide. SARS-CoV-2 is usually detected via real-time reverse-transcription polymerase chain reaction (RT-PCR). However, the increase in specimen load in institutions/hospitals necessitates a simpler detection system. Here, we present an ultra-rapid, real-time RT-PCR assay for SARS-CoV-2 detection using PCR1100 device. Although PCR1100 tests only one specimen at a time, the amplification period is less than 20 min and the sensitivity and specificity match those of conventional real-time RT-PCR performed on large instruments. The method is potentially helpful when daily multiple SARS-CoV-2 testing is needed, for example to confirm virus-free status prior to patient discharge. |
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| AbstractList | The disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in Wuhan, China, in December 2019, has rapidly spread worldwide. SARS-CoV-2 is usually detected via real-time reverse-transcription polymerase chain reaction (RT-PCR). However, the increase in specimen load in institutions/hospitals necessitates a simpler detection system. Here, we present an ultra-rapid, real-time RT-PCR assay for SARS-CoV-2 detection using PCR1100 device. Although PCR1100 tests only one specimen at a time, the amplification period is less than 20 min and the sensitivity and specificity match those of conventional real-time RT-PCR performed on large instruments. The method is potentially helpful when daily multiple SARS-CoV-2 testing is needed, for example to confirm virus-free status prior to patient discharge. The disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in Wuhan, China, in December 2019, has rapidly spread worldwide. SARS-CoV-2 is usually detected via real-time reverse-transcription polymerase chain reaction (RT-PCR). However, the increase in specimen load in institutions/hospitals necessitates a simpler detection system. Here, we present an ultra-rapid, real-time RT-PCR assay for SARS-CoV-2 detection using PCR1100 device. Although PCR1100 tests only one specimen at a time, the amplification period is less than 20 min and the sensitivity and specificity match those of conventional real-time RT-PCR performed on large instruments. The method is potentially helpful when daily multiple SARS-CoV-2 testing is needed, for example to confirm virus-free status prior to patient discharge.The disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in Wuhan, China, in December 2019, has rapidly spread worldwide. SARS-CoV-2 is usually detected via real-time reverse-transcription polymerase chain reaction (RT-PCR). However, the increase in specimen load in institutions/hospitals necessitates a simpler detection system. Here, we present an ultra-rapid, real-time RT-PCR assay for SARS-CoV-2 detection using PCR1100 device. Although PCR1100 tests only one specimen at a time, the amplification period is less than 20 min and the sensitivity and specificity match those of conventional real-time RT-PCR performed on large instruments. The method is potentially helpful when daily multiple SARS-CoV-2 testing is needed, for example to confirm virus-free status prior to patient discharge. |
| Author | Matsuyama, Shutoku Shirato, Kazuya Takeda, Makoto Nao, Naganori Kageyama, Tsutomu |
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| Cites_doi | 10.46405/ejms.v2i2.101 10.2807/1560-7917.ES.2020.25.3.2000045 10.7883/yoken.JJID.2019.400 10.1038/s41586-020-2008-3 10.1016/j.virol.2017.11.012 10.7883/yoken.JJID.2020.061 10.7883/yoken.67.469 10.3201/eid1002.030759 10.7883/yoken.JJID.2020.182 10.1371/journal.pone.0215822 10.1038/s41586-020-2012-7 10.1073/pnas.2002589117 10.1099/vir.0.043117-0 10.1371/journal.pone.0099782 10.1128/JVI.00676-08 |
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| References | 6. Shirato K, Nao N, Kawase M, et al. An ultra-rapid real-time RT-PCR method for detecting human orthopneumovirus using PCR1100. Jpn J Infect Dis. 2020;73:465-8. 4. Shirato K, Nao N, Katano H, et al. Development of genetic diagnostic methods for novel coronavirus 2019 (nCoV-2019) in Japan. Jpn J Infect Dis. 2020;73:304-7. 9. Shirogane Y, Takeda M, Iwasaki M, et al. Efficient multiplication of human metapneumovirus in Vero cells expressing the transmembrane serine protease TMPRSS2. J Virol. 2008;82:8942-6. 14. Emery SL, Erdman DD, Bowen MD, et al. Real-time reverse transcription-polymerase chain reaction assay for SARS-associated coronavirus. Emerg Infect Dis. 2004;10:311-6. 5. Shirato K, Nao N, Matsuyama S, et al. Ultra-rapid real-time RT-PCR method for detecting middle east respiratory syndrome coronavirus using a mobile PCR device, PCR1100. Jpn J Infect Dis. 2020;73:181-6. 12. Owusu M, Annan A, Corman VM, et al. Human coronaviruses associated with upper respiratory tract infections in three rural areas of Ghana. PLoS One. 2014;9:e99782. 8. Shirato K, Kawase M, Watanabe O, et al. Differences in neutralizing antigenicity between laboratory and clinical isolates of HCoV-229E isolated in Japan in 2004-2008 depend on the S1 region sequence of the spike protein. J Gen Virol. 2012;93:1908-17. 15. Corman VM, Landt O, Kaiser M, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020;25:2000045. 10. Kaida A, Kubo H, Takakura K, et al. Associations between co-detected respiratory viruses in children with acute respiratory infections. Jpn J Infect Dis. 2014;67:469-75. 2. Zhou P, Yang XL, Wang XG, et al. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature. 2020;579:270-3. 11. Shirato K, Kawase M, Matsuyama S. Wild-type human coronaviruses prefer cell-surface TMPRSS2 to endosomal cathepsins for cell entry. Virology. 2018;517:9-15. 7. Matsuyama S, Nao N, Shirato K, et al. Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells. Proc Natl Acad Sci U S A. 2020;117:7001-3. 3. WHO. Novel Coronavirus (2019-nCoV) situation reports. Available at <https://www.who.int/emergencies/diseases/novel-coronavirus-2019/situation-reports/>. Accessed June 2, 2020. 1. Wu F, Zhao S, Yu B, et al. A new coronavirus associated with human respiratory disease in China. Nature. 2020;579:265-9. 13. Nao N, Sato K, Yamagishi J, et al. Consensus and variations in cell line specificity among human metapneumovirus strains. PLoS One. 2019;14:e0215822. 11 12 13 14 15 1 2 3 4 5 6 7 8 9 10 |
| References_xml | – reference: 1. Wu F, Zhao S, Yu B, et al. A new coronavirus associated with human respiratory disease in China. Nature. 2020;579:265-9. – reference: 13. Nao N, Sato K, Yamagishi J, et al. Consensus and variations in cell line specificity among human metapneumovirus strains. PLoS One. 2019;14:e0215822. – reference: 15. Corman VM, Landt O, Kaiser M, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020;25:2000045. – reference: 3. WHO. Novel Coronavirus (2019-nCoV) situation reports. Available at <https://www.who.int/emergencies/diseases/novel-coronavirus-2019/situation-reports/>. Accessed June 2, 2020. – reference: 5. Shirato K, Nao N, Matsuyama S, et al. Ultra-rapid real-time RT-PCR method for detecting middle east respiratory syndrome coronavirus using a mobile PCR device, PCR1100. Jpn J Infect Dis. 2020;73:181-6. – reference: 8. Shirato K, Kawase M, Watanabe O, et al. Differences in neutralizing antigenicity between laboratory and clinical isolates of HCoV-229E isolated in Japan in 2004-2008 depend on the S1 region sequence of the spike protein. J Gen Virol. 2012;93:1908-17. – reference: 9. Shirogane Y, Takeda M, Iwasaki M, et al. Efficient multiplication of human metapneumovirus in Vero cells expressing the transmembrane serine protease TMPRSS2. J Virol. 2008;82:8942-6. – reference: 12. Owusu M, Annan A, Corman VM, et al. Human coronaviruses associated with upper respiratory tract infections in three rural areas of Ghana. PLoS One. 2014;9:e99782. – reference: 6. Shirato K, Nao N, Kawase M, et al. An ultra-rapid real-time RT-PCR method for detecting human orthopneumovirus using PCR1100. Jpn J Infect Dis. 2020;73:465-8. – reference: 7. Matsuyama S, Nao N, Shirato K, et al. Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells. Proc Natl Acad Sci U S A. 2020;117:7001-3. – reference: 14. Emery SL, Erdman DD, Bowen MD, et al. Real-time reverse transcription-polymerase chain reaction assay for SARS-associated coronavirus. Emerg Infect Dis. 2004;10:311-6. – reference: 2. Zhou P, Yang XL, Wang XG, et al. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature. 2020;579:270-3. – reference: 11. Shirato K, Kawase M, Matsuyama S. Wild-type human coronaviruses prefer cell-surface TMPRSS2 to endosomal cathepsins for cell entry. Virology. 2018;517:9-15. – reference: 10. Kaida A, Kubo H, Takakura K, et al. Associations between co-detected respiratory viruses in children with acute respiratory infections. Jpn J Infect Dis. 2014;67:469-75. – reference: 4. Shirato K, Nao N, Katano H, et al. Development of genetic diagnostic methods for novel coronavirus 2019 (nCoV-2019) in Japan. Jpn J Infect Dis. 2020;73:304-7. – ident: 3 doi: 10.46405/ejms.v2i2.101 – ident: 15 doi: 10.2807/1560-7917.ES.2020.25.3.2000045 – ident: 5 doi: 10.7883/yoken.JJID.2019.400 – ident: 1 doi: 10.1038/s41586-020-2008-3 – ident: 11 doi: 10.1016/j.virol.2017.11.012 – ident: 4 doi: 10.7883/yoken.JJID.2020.061 – ident: 10 doi: 10.7883/yoken.67.469 – ident: 14 doi: 10.3201/eid1002.030759 – ident: 6 doi: 10.7883/yoken.JJID.2020.182 – ident: 13 doi: 10.1371/journal.pone.0215822 – ident: 2 doi: 10.1038/s41586-020-2012-7 – ident: 7 doi: 10.1073/pnas.2002589117 – ident: 8 doi: 10.1099/vir.0.043117-0 – ident: 12 doi: 10.1371/journal.pone.0099782 – ident: 9 doi: 10.1128/JVI.00676-08 |
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| SubjectTerms | Coronaviruses COVID-19 COVID-19 - virology COVID-19 Testing - instrumentation COVID-19 Testing - methods Humans PCR1100 Polymerase chain reaction Real time Real-Time Polymerase Chain Reaction - instrumentation Real-Time Polymerase Chain Reaction - methods Respiratory diseases Reverse Transcriptase Polymerase Chain Reaction - instrumentation Reverse Transcriptase Polymerase Chain Reaction - methods SARS-CoV-2 - genetics SARS-CoV-2 - isolation & purification Sensitivity and Specificity Severe acute respiratory syndrome coronavirus 2 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ultra-rapid real-time RT-PCR Viral diseases |
| Title | An Ultra-Rapid Real-Time RT-PCR Method Using the PCR1100 to Detect Severe Acute Respiratory Syndrome Coronavirus-2 |
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