Reverse transcriptase loop-mediated isothermal amplification (RT-LAMP)-based diagnosis: A potential alternative to quantitative real-time PCR based detection of the novel SARS-COV-2 virus

被引:9
作者
Haq, Farhan [1 ]
Sharif, Salmaan [2 ]
Khurshid, Adnan [2 ]
Ikram, Aamer [2 ]
Shabbir, Imran [3 ]
Salman, Muhammad [2 ]
Ahad, Abdul [2 ]
Rana, Muhammad Suleman [2 ]
Raja, Aroosha [1 ]
Badar, Nazish [2 ]
Tashkandi, Hanaa [4 ]
Al Amri, Turki [5 ]
Azhar, Esam, I [6 ]
Almuhayawi, Mohammed S. [7 ]
Harakeh, Steve [6 ,8 ]
Malik, Muhammad Faraz Arshad [1 ]
机构
[1] COMSATS Univ Islamabad, Dept Biosci, Islamabad, Pakistan
[2] Natl Inst Hlth, Islamabad, Pakistan
[3] Int Islamic Univ, Dept Biotechnol & Bioinformat, Islamabad, Pakistan
[4] King Abdulaziz Univ KAU, Fac Med FM, Dept Surg, Jeddah, Saudi Arabia
[5] KAU, Family & Community Med Dept, Fac Med Rabigh, Jeddah, Saudi Arabia
[6] KAU, King Fahd Med Res Ctr, Dept Med Lab Sci, Special Infect Agents Unit,Fac Appl Med Sci, Jeddah, Saudi Arabia
[7] KAU, FM, Dept Microbiol & Parasitol, Jeddah, Saudi Arabia
[8] KAU, FM, Yousef Abdul Latif Jameel Sci Chair Prophet Med A, Jeddah, Saudi Arabia
关键词
SARS-CoV-2; COVID-19; RT-LAMP; qRT-PCR; Detection; RNA isolation; RAPID DETECTION; COVID-19;
D O I
10.1016/j.sjbs.2020.10.064
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
The sudden outbreak of the novel Coronavirus infectious disease (COVID-19) resulted in significant challenges to global health systems. One of the primary challenges is rapid, reliable, and accurate detection of the severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) virus among the suspected COVID19-infected individuals. At present, quantitative real-time PCR (qRT-PCR) is a widely used diagnostic method. However, it requires expensive instruments and expertise in the interpretation of results. These constraints reflect the significant need for the development of alternative diagnostic options. This study will validate the use and efficiency of the reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay as a potential alternative for the detection of COVID-19. For this purpose, a cohort of 297 suspected COVID-19 patients was tested using both the RT-LAMP assay and the conventional RT-PCR method. For the RT-LAMP assay, three genes (orf-1ab, N, and S) were identified as the target sites for the detection of COVID-19. Based on a comparative assessment, 117 out of 124 positive COVID19 cases were observed using the RT-LAMP technique with an overall 91.45% sensitivity. Interestingly, where a consensus on 163 individuals free of SARS-Cov-2 was observed, RT-LAMP specificity was 90%. Based on these findings, the robustness of the technique, and the reduced dependency on expensive instrumentation, RT-LAMP-based COVID-19 detection is strongly recommended as a potential alternative assay. (C) 2020 Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license.
引用
收藏
页码:942 / 947
页数:6
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