tetano
Editor, Senior Moderator
BMC Infect Dis
. 2024 Jan 15;24(1):81.
doi: 10.1186/s12879-023-08924-3. Development and clinical application of loop-mediated isothermal amplification combined with lateral flow assay for rapid diagnosis of SARS-CoV-2
Jin Tang[SUP] #[/SUP][SUP] 1 [/SUP], Jie Zhu[SUP] #[/SUP][SUP] 2 3 [/SUP], Jie Wang[SUP] #[/SUP][SUP] 4 5 [/SUP], Haiyong Qian[SUP] 6 [/SUP], Zengxin Liu[SUP] 7 [/SUP], Ru Wang[SUP] 7 [/SUP], Qingqing Cai[SUP] 7 [/SUP], Yuan Fang[SUP] 8 [/SUP], Weifeng Huang[SUP] 9 [/SUP]
Affiliations
Background: The diagnostic assay leveraging multiple reverse transcription loop-mediated isothermal amplification (RT-LAMP) could meet the requirements for rapid nucleic acid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
Methods: The devised assay merged the lateral flow assay with the RT-LAMP technology and designed specific primers for the simultaneous detection of the target and human-derived internal reference genes within a single reaction. An inquiry into the assay's limit of detection (LOD), sensitivity, and specificity was carried out. The effectiveness of this assay was validated using 498 clinical specimens.
Results: This LOD of the assay was determined to be 500 copies/mL, and there was no observed cross-reaction with other respiratory pathogens. The detection results derived from clinical specimens showed substantial concordance with those from real-time reverse transcription-polymerase chain reaction (RT-qPCR) (Cohen's kappa, 0.876; 95% CI: 0.833-0.919; p<0.005). The diagnostic sensitivity and specificity were 87.1% and 100%, respectively.
Conclusion: The RT-LAMP assay, paired with a straightforward and disposable lateral immunochromatographic strip, achieves visual detection of dual targets for SARS-CoV-2 immediatly. Moreover, the entire procedure abstains from nucleic acids extraction. The samples are lysed at room temperature and subsequently proceed directly to the RT-LAMP reaction, which can be executed within 30 minutes at a constant temperature of 60-65°C. Then, the RT-LAMP amplification products are visualized using colloidal gold test strips.
Trial registration: This study was registered at the Chinese Clinical Trial Registry (Registration number: ChiCTR2200060495, Date of registration 2022-06-03).
Keywords: COVID-19; Diagnostics; Isothermal amplification; Molecular testing; RT-LAMP; SARS-CoV-2.
. 2024 Jan 15;24(1):81.
doi: 10.1186/s12879-023-08924-3. Development and clinical application of loop-mediated isothermal amplification combined with lateral flow assay for rapid diagnosis of SARS-CoV-2
Jin Tang[SUP] #[/SUP][SUP] 1 [/SUP], Jie Zhu[SUP] #[/SUP][SUP] 2 3 [/SUP], Jie Wang[SUP] #[/SUP][SUP] 4 5 [/SUP], Haiyong Qian[SUP] 6 [/SUP], Zengxin Liu[SUP] 7 [/SUP], Ru Wang[SUP] 7 [/SUP], Qingqing Cai[SUP] 7 [/SUP], Yuan Fang[SUP] 8 [/SUP], Weifeng Huang[SUP] 9 [/SUP]
Affiliations
- PMID: 38225546
- DOI: 10.1186/s12879-023-08924-3
Background: The diagnostic assay leveraging multiple reverse transcription loop-mediated isothermal amplification (RT-LAMP) could meet the requirements for rapid nucleic acid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
Methods: The devised assay merged the lateral flow assay with the RT-LAMP technology and designed specific primers for the simultaneous detection of the target and human-derived internal reference genes within a single reaction. An inquiry into the assay's limit of detection (LOD), sensitivity, and specificity was carried out. The effectiveness of this assay was validated using 498 clinical specimens.
Results: This LOD of the assay was determined to be 500 copies/mL, and there was no observed cross-reaction with other respiratory pathogens. The detection results derived from clinical specimens showed substantial concordance with those from real-time reverse transcription-polymerase chain reaction (RT-qPCR) (Cohen's kappa, 0.876; 95% CI: 0.833-0.919; p<0.005). The diagnostic sensitivity and specificity were 87.1% and 100%, respectively.
Conclusion: The RT-LAMP assay, paired with a straightforward and disposable lateral immunochromatographic strip, achieves visual detection of dual targets for SARS-CoV-2 immediatly. Moreover, the entire procedure abstains from nucleic acids extraction. The samples are lysed at room temperature and subsequently proceed directly to the RT-LAMP reaction, which can be executed within 30 minutes at a constant temperature of 60-65°C. Then, the RT-LAMP amplification products are visualized using colloidal gold test strips.
Trial registration: This study was registered at the Chinese Clinical Trial Registry (Registration number: ChiCTR2200060495, Date of registration 2022-06-03).
Keywords: COVID-19; Diagnostics; Isothermal amplification; Molecular testing; RT-LAMP; SARS-CoV-2.