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

被引:6
|
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
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