著者
Kazuya Shirato Naganori Nao Harutaka Katano Ikuyo Takayama Shinji Saito Fumihiro Kato Hiroshi Katoh Masafumi Sakata Yuichiro Nakatsu Yoshio Mori Tsutomu Kageyama Shutoku Matsuyama Makoto Takeda
出版者
National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee
雑誌
Japanese Journal of Infectious Diseases (ISSN:13446304)
巻号頁・発行日
pp.JJID.2020.061, (Released:2020-02-18)
参考文献数
7
被引用文献数
64

At the end of 2019, pneumonia caused by novel coronavirus 2019 (nCoV) emerged in Wuhan city, China. Many airline travelers moved between Wuhan and Japan at that time, suggesting that Japan is at high risk of invasion by the virus. Diagnostic systems for 2019-nCoV were developed with urgency. Two nested RT–PCR assays and two real-time RT–PCR assays were adapted to local Japanese conditions. As of 8 February 2020, the assays developed have successfully detected 25 positive cases of infection in Japan.
著者
Kiyoko Okamaoto Kazuya Shirato NagaNaganori Nao Shinji Saito Tsutomu Kageyama Hideki Hasegawa Tadaki Suzuki Shutoku Matsuyama Makoto Takeda
出版者
National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee
雑誌
Japanese Journal of Infectious Diseases (ISSN:13446304)
巻号頁・発行日
pp.JJID.2020.108, (Released:2020-04-30)
参考文献数
11
被引用文献数
13 18

The COVID-19 outbreak caused by SARS-CoV-2 in Wuhan (China) in December 2019 is currently spreading rapidly and globally. We recently reported a laboratory diagnostic protocol for SARS-CoV-2 based on real-time RT-PCR assays using two primer sets, N and N2. On 30–31 January 2020, the protocol and reagents for these assays were distributed to local public health institutes and quarantine depots in Japan, and nationwide, SARS-CoV-2 diagnostic testing was started. For further validation, the assays were compared with the commercially available kits using SARS CoV-2 viral RNA and the clinical specimens obtained from COVID19-suspected individuals. The LightMix Modular SARS and Wuhan CoV E-gene (LN S&W-E) assay was highly sensitive for SARS-CoV-2, as was the N2 set, and both assays had perfectly consistent results with the clinical specimens. While the LM S&W-E set targets the highly conserved region of the E gene in SARS-CoV and SARS-CoV-2, the N2 set was designed to target the unique region in the SARS-CoV-2 N gene. Thus, the N2 set has high specificity and sensitivity for SARS-CoV-2 detection. These indicate that the protocol using the N and N2 sets is comparable to commercially available kits and is reliable for the laboratory diagnosis of COVID-19.
著者
Kazuya Shirato Naganori Nao Shutoku Matsuyama Makoto Takeda Tsutomu Kageyama
出版者
National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee
雑誌
Japanese Journal of Infectious Diseases (ISSN:13446304)
巻号頁・発行日
pp.JJID.2020.324, (Released:2020-06-30)
参考文献数
15
被引用文献数
1

The disease caused by severe acute respiratory syndrome coronavirus (SARS-CoV-2) in Wuhan, China, in December 2019 is currently spreading rapidly worldwide. SARS-CoV-2 is usually detected via real-time RT-PCR. However, as institutions/hospitals deal with increasing numbers of specimens, a simpler detection system is required. Here, we present an ultra-rapid, real-time RT-PCR assay for SARS-CoV-2 using the PCR1100 device. Although this tests only one specimen at any one time, the amplification period is <20 min, with maintenance of the sensitivity and specificity of conventional real-time RT-PCR performed using large instruments. The method will be very helpful if SARS-CoV-2 testing is required a few times daily, for example to confirm virus-free status prior to discharge.
著者
Kazuya Shirato Naganori Nao Shutoku Matsuyama Tsutomu Kageyama
出版者
National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee
雑誌
Japanese Journal of Infectious Diseases (ISSN:13446304)
巻号頁・発行日
pp.JJID.2019.400, (Released:2019-12-25)
参考文献数
21
被引用文献数
4

Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) is usually diagnosed through highly sensitive and specific genetic tests such as real-time reverse transcription polymerase chain reaction (RT-PCR). Currently, two real-time RT-PCR assays targeting the upE and ORF1a regions of the MERS-CoV genome are widely used and are the standard assays recommended by the World Health Organization (WHO). The MERS outbreaks to date suggest that rapid diagnosis and subsequent isolation of infected patients, particularly superspreaders, are critical for containment. However, conventional real-time RT-PCR assays require large laboratory instruments, and amplification takes approximately 2 h. These are disadvantages for rapid diagnosis. Here, an ultra-rapid real-time RT-PCR test was established: a multiplex assay for upE and ORF1a running on the mobile PCR1100 device. As few as five copies of MERS-CoV RNA can be detected within 20 min using the WHO standard assays with similar sensitivity and specificity to those of a conventional real-time PCR instrument such as the LightCyler, enabling timely intervention to control MERS-CoV infection.