Ultrafast and absolute quantification of SARS-CoV-2 on food using hydrogel RT-LAMP without pre-lysis

Isothermal amplification-based microfluidic devices for detecting foodborne pathogens: a review

“…An ultrafast hydrogel-mediated RT-LAMP point-of-care testing system was developed to screen Critical Review Analytical Methods SARS-CoV2 on cold chain fruits. 44 In this study, a hydrogel was employed to provide an environment for the RT-LAMP reaction, which improved amplication efficiency and analysis time. An advanced microuidic device was developed to detect SARS-CoV2 in wastewater.…”

Ultrasensitive Hybridization Chain Reaction-Assisted Multisite Exonuclease III Amplification Strategy Combined with a Direct Quantitative Fluorescence Lateral Flow Technique for Multiple Bacterial 16S rRNA Detection

“…Current RNA detection methods such as northern blotting, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and RT-loop-mediated isothermal amplification (RT-LAMP) have been popularly employed at different occasions, − but they also exhibit a few limitations. Northern blotting is a classic method, but the sensitivity remains to be improved .…”

Hydrogel Digital LAMP with Suppressed Nonspecific Amplification for Rapid Diagnostics of Fungal Disease in Fresh Fruits

“…Recently, molecular techniques have been extensively applied for fungal species identification, such as polymerase chain reaction (PCR) and real-time PCR (qPCR) using specific primers to amplify target genomic segments. , Digital PCR is a single-molecule detection technique that dispenses reaction solution into thousands of microwells, followed by single nucleic acid amplification and end-point digital counting, which is faster and more sensitive than conventional semiquantitative methods. , However, PCR relies on the costly specialized equipment and thermal cycle with long amplification time . Alternatively, loop-mediated isothermal amplification (LAMP) has become an effective assay for point-of-care test. , LAMP can be operated at a constant temperature with high accuracy and high sensitivity. − Nevertheless, nonspecific amplification is an inevitable process in the rapid nucleic acid test, adversely affecting the diagnostic accuracy and sensitivity of fungal diseases with false positives.…”

“…Although probes or molecule beacons can be applied to detect specific signals, nonspecific amplification cannot be eliminated in the assay, which can compete with the specific positive amplification. , Recently, various strategies have been used to avoid nonspecific amplification, like adding nanomaterials, single-strand binding proteins, or chemical additives (like pullulan or dimethyl sulfoxide) into the assay. − However, these methods are costly and require the introduction of other reagents. Meanwhile, with the suppression of nonspecific amplification, the specific positive amplification would also be affected, limiting their sensitivity and detection speed. , The hydrogel system offers an excellent platform for nucleic acid analysis. , The movement of nucleic acid is limited in the hydrogel, while the diffusion of small molecules is allowed. ,, The cross-linking networks of hydrogels could also be applied for single nucleic acid extraction and single-cell analysis. − However, its potential for diagnosing fungal diseases has not been explored, which requires high specificity and accuracy without false positive results.…”

“…39,40 The movement of nucleic acid is limited in the hydrogel, while the diffusion of small molecules is allowed. 22,23,41 The cross-linking networks of hydrogels could also be applied for single nucleic acid extraction and single-cell analysis. 42−45 However, its potential for diagnosing fungal diseases has not been explored, which requires high specificity and accuracy without false positive results.…”