Comparison of RT-LAMP and RT-qPCR assays for detecting SARS-CoV-2 in the extracted RNA and direct swab samples

被引:7
作者
Pourakbari, Ramin [1 ,2 ]
Gholami, Mohammad [1 ]
Shakerimoghaddam, Ali [1 ,2 ]
Khiavi, Farhad Motavalli [1 ,2 ]
Mohammadimehr, Mojgan [1 ]
Khomartash, Mehdi Shakouri [1 ,2 ]
机构
[1] Aja Univ Med Sci, Infect Dis Res Ctr, Tehran, Iran
[2] Aja Univ Med Sci, Med Biotechnol Res Ctr, Etemadzadeh St, Tehran, Iran
关键词
SARS-CoV-2; RT-LAMP; RT-qPCR; Direct assay;
D O I
10.1016/j.jviromet.2023.114871
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in infected patients is critical for infection control. Loop-mediated isothermal amplification (LAMP) has been demonstrated to be a rapid, simple, reliable, cost-effective and sensitive method to detect SARS-CoV-2 in a variety of samples in considerably less time than Real-Time PCR. In this study, we developed and optimized a rapid detection method for SARSCoV-2 based on RT-LAMP method utilizing a specific primer set targeting the ORF1a gene and then examined its sensitivity and efficiency using a serially diluted viral RNA sample with a known concentration. Furthermore, the sensitivity of the RT-LAMP to detect SARS-CoV-2 in direct swab samples with varying Ct values were compared to a commercial molecular RT-qPCR based detection kit. According to our findings the optimal incubation time for RT-LAMP assay was 45 min. There was a complete agreement between RT-LAMP and RT-qPCR in diagnosing the viral genome in the diluted extracted RNA sample. However, it had a lower sensitivity (71%) to detect the viral genome in direct swab samples compared to RT-qPCR. In conclusion, due to its simplicity, rapidness, sensitivity, and specificity, RT-LAMP has tremendous potential as a point-of-care tool; nevertheless, more research is needed to utilize it for detecting SARS-CoV-2, particularly in direct swab samples.
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页数:5
相关论文
共 22 条
[1]  
Abdullahi U.F., 2015, INDIAN J SCI TECHNOL, V8, P1
[2]   Development of loop-mediated isothermal amplification (LAMP) assays using five primers reduces the false-positive rate in COVID-19 diagnosis [J].
Alhamid, Galyah ;
Tombuloglu, Huseyin ;
Al-Suhaimi, Ebtesam .
SCIENTIFIC REPORTS, 2023, 13 (01)
[3]   Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load [J].
Aoki, Mateus Nobrega ;
Coelho, Bruna de Oliveira ;
Bentim Goes, Luiz Gustavo ;
Minoprio, Paola ;
Durigon, Edison Luiz ;
Morello, Luis Gustavo ;
Marchini, Fabricio Klerynton ;
Riediger, Irina Natassja ;
Debur, Maria do Carmo ;
Nakaya, Helder I. ;
Blanes, Lucas .
SCIENTIFIC REPORTS, 2021, 11 (01)
[4]  
Buck M.D., 2021, Wellcome Open Res, V6
[5]   A Rapid, Simple, Inexpensive, and Mobile Colorimetric Assay COVID-19-LAMP for Mass On-Site Screening of COVID-19 [J].
Chow, Franklin Wang-Ngai ;
Chan, Tony Tat-Yin ;
Tam, Anthony Raymond ;
Zhao, Suhui ;
Yao, Weiming ;
Fung, Joshua ;
Cheng, Flora Ka-Kei ;
Lo, George Chi-Shing ;
Chu, Stella ;
Aw-Yong, Kam Leng ;
Tang, James Yat-Man ;
Tsang, Chi-Ching ;
Luk, Hayes Kam-Hei ;
Wong, Antonio Cheuk-Pui ;
Li, Kenneth Sze-Ming ;
Zhu, Longchao ;
He, Zirong ;
Tam, Emily Wan Ting ;
Chung, Tom Wai-Hin ;
Wong, Sally Cheuk Ying ;
Que, Tak-Lun ;
Fung, Kitty Sau-Chun ;
Lung, David Christopher ;
Wu, Alan Ka-Lun ;
Hung, Ivan Fan-Ngai ;
Woo, Patrick Chiu-Yat ;
Lau, Susanna Kar-Pui .
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2020, 21 (15) :1-10
[6]  
Cisneros-Villanueva M., 2022, MedRxiv, V2022, P15
[7]   Rapid and Reliable Detection of SARS-CoV-2 Using Direct RT-LAMP [J].
El-Kafrawy, Sherif A. ;
El-Daly, Mai M. ;
Hassan, Ahmed M. ;
Harakeh, Steve M. ;
Alandijany, Thamir A. ;
Azhar, Esam, I .
DIAGNOSTICS, 2022, 12 (04)
[8]   Shining a light on LAMP assays-A comparison of LAMP visualization methods including the novel use of berberine [J].
Fischbach, Jens ;
Xander, Nina Carolin ;
Frohme, Marcus ;
Gloekler, Joern Felix .
BIOTECHNIQUES, 2015, 58 (04) :189-194
[9]   Developing RT-LAMP assays for rapid diagnosis of SARS-CoV-2 in saliva [J].
Huang, Xin ;
Tang, Gongyu ;
Ismail, Nahed ;
Wang, Xiaowei .
EBIOMEDICINE, 2022, 75
[10]   Ct values as a diagnostic tool for monitoring SARS-CoV-2 viral load using the QIAstat-Dx® Respiratory SARS-CoV-2 Panel [J].
Juanola-Falgarona, Marti ;
Penarrubia, Luis ;
Jimenez-Guzman, Sara ;
Porco, Roberto ;
Congost-Teixidor, Claudia ;
Varo-Velazquez, Marta ;
Rao, Sonia N. ;
Pueyo, Gemma ;
Manissero, Davide ;
Pareja, Josep .
INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES, 2022, 122 :930-935