Loop-mediated isothermal amplification for rapid detection of Acute viral necrobiotic virus in scallop Chlamys farreri

Development and Application of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Fusarium Oxysporum f. Sp. lycopersici

“…In this context, a sky blue colour pattern implies the existence of the reference virus, whereas a violet colour change is observed when the control(s) are taken into consideration. , diluted to 1:10 with 6× loading buffer (TaKara, Dalian, China), was added to each reaction as described previously [23,41,43]. Remarkably, concerning negative reaction, the original orange colour could be observed.…”

“…Remarkably, concerning negative reaction, the original orange colour could be observed. SYBR Green I: 1 μl of SYBR Green I (Invitrogen, Sydney, Australia) diluted to 1:10 with 6× loading buffer Was separately added to each reaction as described previously [23,44]. Remarkably, concerning negative reaction, the original orange colour could be observed.…”

“…LAMP assay can also amplify nucleic acid under isothermal condition in the range of 60 to 65°C, all turbidity-and fluorescent-based detections, as well as agarose gel electrophoresis system are applied to visualize suspicious samples [15]. LAMP-positive amplicons confirmed by adding a number of fluorescent dsDNA intercalating dye including ethidium bromide [18,19], SYBR Green I [20] and propidium iodide [21] after the reaction is completed or metal indicators such as calcein [22], GeneFinder TM [23], hydroxynaphthol blue (HNB) [24][25][26] and magnesium pyrophosphate [14,18,27] prior to the reaction, allowing observation with the naked eye [28,29]. LAMP assay was widely applied for detection of many important infectious diseases in human and animals [30].…”

Colorimetric Immunocapture Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid Detection of the Potato virus Y

“…One set of primer (Table 1), that is backward and forward primers, was used for the PCR amplification of the cDNA. Amplification was performed with the following PCR profile: 94°C for 3 min; 35 cycles of 1 min at 94°C, 1 min at 54°C, 1 min at 72°C and 10 to 1:10 with 6× loading buffer (TaKara, Dalian, China), was added to each reaction as described previously [29,30]. Remarkably, concerning negative reaction, the original orange color could be observed.…”

“…Notably, despite a few number of studies about immunocapture reverse transcription loopmediated isothermal amplification (IC-RT-LAMP) [17,24] because the technique has not been yet introduced for detection of PVY, an attempt was accordingly made to optimize a new protocol of it to save time and remove RNA extraction. As the second purpose, since the existence of one-step IC-RT-LAMP positive amplicons has been proved to be confirmed by adding a number of fluorescent dsDNA intercalating dye including ethidum bromide [25], SYBR Green [26] and propidium iodide [27] after the reaction is completed or metal indicators such as calcein [28], Gene-Finder TM [29,30], hydroxyl naphthol blue [31][32][33] and magnesium pyrophosphate [17,18] prior to the reaction, allowing observation with the naked eye, here, six different visualization systems were consequently employed to assess the colour stability as well as safety feature of each one in a viral detection procedure of PVY.…”