Nucleic acid visualization assay for Middle East Respiratory Syndrome Coronavirus (MERS-CoV) by targeting the UpE and N gene
| العنوان: | Nucleic acid visualization assay for Middle East Respiratory Syndrome Coronavirus (MERS-CoV) by targeting the UpE and N gene |
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| المؤلفون: | Tiecheng Wang, Pei Huang, Entao Li, Na Feng, Yongkun Zhao, Songtao Yang, Yumeng Song, Hongli Jin, Wujian Li, Hongbin He, Hang Chi, Cuicui Jiao, Feihu Yan, Jianzhong Wang, Qi Wang, Hualei Wang, Ruo Mo, Jinhao Bi, Qiuxue Han, Aimin Ma, Hongmei Wang, Yuwei Gao, Xianzhu Xia |
| المصدر: | PLoS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, Vol 15, Iss 3, p e0009227 (2021) |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | RNA viruses, Viral Diseases, Pulmonology, Coronaviruses, viruses, RC955-962, Recombinase Polymerase Amplification, Viral Nonstructural Proteins, medicine.disease_cause, Pathology and Laboratory Medicine, Biochemistry, Medical Conditions, Camels, Arctic medicine. Tropical medicine, Nucleic Acids, Medicine and Health Sciences, Respiratory system, Coronavirus, Virus Testing, Mammals, virus diseases, Eukaryota, Infectious Diseases, Molecular Diagnostic Techniques, Medical Microbiology, Viral Pathogens, Viruses, Vertebrates, Middle East Respiratory Syndrome Coronavirus, RNA, Viral, RNA extraction, Public aspects of medicine, RA1-1270, Pathogens, Coronavirus Infections, Research Article, Middle East respiratory syndrome coronavirus, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Saudi Arabia, Biology, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Microbiology, Respiratory Disorders, Extraction techniques, Diagnostic Medicine, medicine, Humans, Animals, Gene, Microbial Pathogens, SARS, Biology and life sciences, business.industry, Clinical Laboratory Techniques, Public Health, Environmental and Occupational Health, Organisms, Outbreak, Covid 19, Reverse Transcription, Virology, Reverse transcriptase, respiratory tract diseases, Research and analysis methods, Respiratory Infections, Amniotes, Nucleic acid, business, Zoology |
| الوصف: | Since its first emergence in 2012, cases of infection with Middle East respiratory syndrome coronavirus (MERS-CoV) have continued to occur. At the end of January 2020, 2519 laboratory confirmed cases with a case-fatality rate of 34.3% have been reported. Approximately 84% of human cases have been reported in the tropical region of Saudi Arabia. The emergence of MERS-CoV has highlighted need for a rapid and accurate assay to triage patients with a suspected infection in a timely manner because of the lack of an approved vaccine or an effective treatment for MERS-CoV to prevent and control potential outbreaks. In this study, we present two rapid and visual nucleic acid assays that target the MERS-CoV UpE and N genes as a panel that combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). This test panel was designed to improve the diagnostic accuracy through dual-target screening after referencing laboratory testing guidance for MERS-CoV. The limit of detection was 1.2×101 copies/μl viral RNA for the UpE assay and 1.2 copies/μl viral RNA for the N assay, with almost consistent with the sensitivity of the RT-qPCR assays. The two assays exhibited no cross-reactivity with multiple CoVs, including the bat severe acute respiratory syndrome related coronavirus (SARSr-CoV), the bat coronavirus HKU4, and the human coronaviruses 229E, OC43, HKU1 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Furthermore, the panel does not require sophisticated equipment and provides rapid detection within 30 min. This panel displays good sensitivity and specificity and may be useful to rapidly detect MERS-CoV early during an outbreak and for disease surveillance. Author summary Symptoms of MERS are atypical, including fever, cough, shortness of breath and occasionally pneumonia and gastrointestinal symptoms. As a result, it is difficult to distinguish from other respiratory pathogens based on the clinical symptoms. MERS-CoV is routinely diagnosed using real-time RT-PCR. Real-time RT-PCR has great advantages in sensitivity and specificity compared with other molecular techniques; nevertheless, it is only suitable for well-equipped or central laboratories due to the demand for sophisticated alternating temperature and fluorescent capture instruments. Here, we developed and evaluated the MERS-CoV RT-RPA-VF assay. Compared to the gold standard real-time RT-PCR, the former was less time consuming and had lower equipment requirements. Thus, this panel may be preferred to rapidly detect MERS-CoV in low-resource settings, even as a point-of-care approach for the timely prevention and control of a pandemic. |
| تدمد: | 1935-2735 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3c56f7bed6e6f171670f0f70377fabb1Test https://pubmed.ncbi.nlm.nih.gov/33647020Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....3c56f7bed6e6f171670f0f70377fabb1 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19352735 |
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