A comparison of single nucleotide primer extension with mispairing PCR-RFLP in detecting a point mutation

Lin, Fu-Hai; Lin, Ruth

doi:10.1016/0006-291x(92)92332-r

Cited by 14 publications

(1 citation statement)

References 7 publications

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“…For the purpose of viral RNA detection, new techniques for rapid isothermal amplification, which is pivotal in rapid point-of-care detection, have been realized using, for example, recombinase polymerase amplification (RPA) [ 13 – 15 ], rolling circle amplification (RCA) [ 16 , 17 ], and loop-mediated isothermal amplification (LAMP) [ 18 , 19 ]. Nevertheless, some of isothermal amplification-based point-of-care detection systems have a limitation because they operate with low selectivity when compared with the RT-PCR method for the diagnosis of viral RNA [ 20 – 22 ]; the RT-PCR method itself also exhibits the same selectivity problem, because of mispairing or self-pairing of the primer during the reverse transcription targeting viral RNA, but recently this problem has been overcome in several ways [ 23 , 24 ]. Point-of-care detection still requires the issue of selectivity to be addressed when using isothermal amplification methods.…”

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confidence: 99%

Dual-site ligation-assisted loop-mediated isothermal amplification (dLig-LAMP) for colorimetric and point-of-care determination of real SARS-CoV-2

2022

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A probing system has been developed based on dual-site ligation-assisted loop-mediated isothermal amplification (dLig-LAMP) for the selective colorimetric detection of SARS-CoV-2. This approach can induce false-positive and -negative detection in real clinical samples; dLig-LAMP operates with improved selectivity. Unlike RT-LAMP, the selectivity of dLig-LAMP is determined in both the ligation and primer binding steps, not in the reverse transcription step. With this selective system in hand, we developed a colorimetric signaling system for point-of-care detection. We also developed a colorimetric probe for sensing pyrophosphate, which arises as a side product during the LAMP DNA amplification. Thus, dLig-LAMP appears to be an alternative method for improving the selectivity problems associated with reverse transcription. In addition, combining dLig-LAMP with colorimetric pyrophosphate probing allows point-of-care detection of SARS-CoV-2 within 1 h with high selectivity. Graphical abstract Supplementary Information The online version contains supplementary material available at 10.1007/s00604-022-05293-7.

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