Rapid, sensitive, and specific detection of SARS-CoV-2 in nasopharyngeal swab samples of suspected patients using a novel one-step loop-mediated isothermal amplification (one-step LAMP) technique

BMC Microbiology

article

Khanizadeh, Sayyad¹  Malekshahi, Asra²  Hanifehpour, Hooman³  Birjandi, Mehdi⁴  Fallahi, Shirzad¹ 

[1] Hepatitis Research Center, School of Medicine, Lorestan University of Medical Sciences;Department of Virology, School of Medicine, Lorestan University of Medical Sciences;Department of Microbiology, Cancer Biomedical Research Center;Department of Biostatistics and Epidemiology, School of Health and Nutrition, Lorestan University of Medical Sciences;Department of Parasitology and Mycology, School of Medicine, Lorestan University of Medical Sciences

关键词: Rapid;    Sensitive;    Detection;    SARS-CoV-2;    One-step LAMP;    One-step RT-qPCR;   

DOI  :  10.1186/s12866-023-02806-z

学科分类：放射科、核医学、医学影像

来源: BioMed Central

【 摘 要 】

In the absence of effective antiviral drugs or vaccines, early and accurate detection of SARS-CoV-2 infection is essential to the COVID-19 pandemic. This study developed and evaluated a novel rapid One-Step LAMP assay to directly detect the SARS-CoV-2 RNA from nasopharyngeal (NP) swab samples of patients with suspected SARS-CoV-2 infection living in deprived areas in comparison to One-Step Real-time PCR. Two hundred fifty-four NP swab samples from patients suspected of COVID-19 infection living in deprived western areas of Iran were tested by TaqMan One-Step RT-qPCR and fast One-Step LAMP assays. Tenfold serial dilutions of SARS-CoV-2 RNA standard strain where the viral copy number in each dilution was previously determined using the qPCR and various templates were used to investigate the analytical sensitivity and specificity of the One-Step LAMP assay in triplicate. Also, the efficacy and reliability of the method compared to TaqMan One-Step RT-qPCR were evaluated using SARS-CoV-2 positive and negative clinical samples. The results of the One-Step RT-qPCR and One-Step LAMP tests were positive in 131 (51.6%) and 127 (50%) participants, respectively. Based on Cohen’s kappa coefficient (κ), the agreement between the two tests was 97%, which was statistically significant (P < 0.001). The detection limit for the One-Step LAMP assay was 1 × 101 copies of standard SARS-CoV-2 RNA per reaction in less than an hour in triplicates. Negative results in all samples with non-SARS-CoV-2 templates represent 100% specificity. The results showed that the One-Step LAMP assay is an efficient consistent technique for detecting SARS-CoV-2 among suspected individuals due to its simplicity, speed, low cost, sensitivity, and specificity. Therefore, it has great potential as a useful diagnostic tool for disease epidemic control, timely treatment, and public health protection, especially in poor and underdeveloped countries.

【 授权许可】

CC BY|CC0