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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 May 22; 73(3):181-186.JJ

Abstract

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 these 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 disadvantages limit rapid diagnosis. Here, an ultra-rapid real-time RT-PCR test was established comprising a multiplex assay for upE and ORF1a running on a mobile PCR1100 device. As few as five copies of the MERS-CoV RNA can be detected within 20 min using the standard WHO assays in the mobile PCR device, with the sensitivity and specificity being similar to those of a conventional real-time PCR instrument such as the LightCyler, thereby enabling timely intervention to control MERS-CoV infection.

Authors+Show Affiliations

Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Diseases.Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Diseases.Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Diseases.Influenza Virus Research Center, National Institute of Infectious Diseases.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31875608

Citation

Kazuya, Shirato, et al. "Ultra-Rapid Real-Time RT-PCR Method for Detecting Middle East Respiratory Syndrome Coronavirus Using a Mobile PCR Device, PCR1100." Japanese Journal of Infectious Diseases, vol. 73, no. 3, 2020, pp. 181-186.
Kazuya S, 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(3):181-186.
Kazuya, S., Nao, N., Matsuyama, S., & Kageyama, T. (2020). Ultra-Rapid Real-Time RT-PCR Method for Detecting Middle East Respiratory Syndrome Coronavirus Using a Mobile PCR Device, PCR1100. Japanese Journal of Infectious Diseases, 73(3), 181-186. https://doi.org/10.7883/yoken.JJID.2019.400
Kazuya 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 May 22;73(3):181-186. PubMed PMID: 31875608.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Ultra-Rapid Real-Time RT-PCR Method for Detecting Middle East Respiratory Syndrome Coronavirus Using a Mobile PCR Device, PCR1100. AU - Kazuya,Shirato, AU - Nao,Naganori, AU - Matsuyama,Shutoku, AU - Kageyama,Tsutomu, Y1 - 2019/12/25/ PY - 2019/12/26/pubmed PY - 2020/9/20/medline PY - 2019/12/26/entrez KW - MERS coronavirus (MERS-CoV) KW - Middle East Respiratory Syndrome (MERS) KW - PCR1100 KW - ultra-rapid real-time RT-PCR SP - 181 EP - 186 JF - Japanese journal of infectious diseases JO - Jpn J Infect Dis VL - 73 IS - 3 N2 - 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 these 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 disadvantages limit rapid diagnosis. Here, an ultra-rapid real-time RT-PCR test was established comprising a multiplex assay for upE and ORF1a running on a mobile PCR1100 device. As few as five copies of the MERS-CoV RNA can be detected within 20 min using the standard WHO assays in the mobile PCR device, with the sensitivity and specificity being similar to those of a conventional real-time PCR instrument such as the LightCyler, thereby enabling timely intervention to control MERS-CoV infection. SN - 1884-2836 UR - https://www.unboundmedicine.com/medline/citation/31875608/Ultra_Rapid_Real_Time_RT_PCR_Method_for_Detecting_Middle_East_Respiratory_Syndrome_Coronavirus_Using_a_Mobile_PCR_Device_PCR1100_ L2 - https://dx.doi.org/10.7883/yoken.JJID.2019.400 DB - PRIME DP - Unbound Medicine ER -