Journal: International Journal of Molecular Sciences

Article Title: Loop-Mediated Isothermal Amplification Assay for the Detection of Citrus Canker Causing Bacterial Variant, Xanthomonas citri pv. citri A w Strain

doi: 10.3390/ijms252111590

Figure Lengend Snippet: Determination of the detection limit of LAMP-A w assay using a serially diluted recombinant plasmid containing the LAMP target (F3-B3 PCR amplicon). ( A ) Real-time PCR (qPCR) amplification curve conducted with LAMP F3/B3 primer set using a serially diluted recombinant plasmid as a template. ( B ) Standard curve generated with the qPCR results in ( A ). The table on the right shows the average qPCR Ct value of triplicates in ( A ) and the Log 10 copy number of the target molecule of each sample that was calculated using a linear regression equation of the standard curve; y = −3.8721x + 41.161 (R 2 = 0.9978) where y and x denote Ct and the Log 10 target copy number, respectively. ( C ) The amplification curve of the LAMP-A w assay was conducted with the serially diluted recombinant plasmid DNA that was used in ( A ). The relative fluorescence unit (RFU) of the LAMP reaction in ( C ) was measured every 30 s (i.e., one LAMP cycle = 30 s). The sample IDs are indicated in the figure. The threshold lines for qPCR and LAMP-A w are indicated in the amplification graphs in ( A ) and ( C ), respectively.

Article Snippet: In order to examine the field applicability of the LAMP-A w assay, the LAMP-A w reaction was conducted with crude DNA extracts prepared with 0.8% NaOH or AMP1 buffer (Agdia) following the manufacturer’s instructions, where the sample boiling step is not required for the crude DNA extract preparation.

Techniques: Recombinant, Plasmid Preparation, Amplification, Real-time Polymerase Chain Reaction, Generated, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Loop-Mediated Isothermal Amplification Assay for the Detection of Citrus Canker Causing Bacterial Variant, Xanthomonas citri pv. citri A w Strain

doi: 10.3390/ijms252111590

Figure Lengend Snippet: The specificity of the LAMP-A w assay using ( A ) DNA fractions prepared from Candidatus Liberibacter asiaticus (CLas)-positive and canker-negative leaf samples collected in the field and ( B ) two Xcc A positives (Xcc A(+)) and one Xcc A w positive DNA sample (Xcc A w (+)). The non-template control (NTC) is indicated in the figure. The threshold line was indicated in the graph. The RFU of the LAMP-A w reaction was monitored every 30 s (i.e., one LAMP cycle = 30 s).

Article Snippet: In order to examine the field applicability of the LAMP-A w assay, the LAMP-A w reaction was conducted with crude DNA extracts prepared with 0.8% NaOH or AMP1 buffer (Agdia) following the manufacturer’s instructions, where the sample boiling step is not required for the crude DNA extract preparation.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Loop-Mediated Isothermal Amplification Assay for the Detection of Citrus Canker Causing Bacterial Variant, Xanthomonas citri pv. citri A w Strain

doi: 10.3390/ijms252111590

Figure Lengend Snippet: Evaluation of the specificity of LAMP-A w assay coupled with lateral flow immunoassay. Sample IDs are shown on top. Samples 77 and 78 were prepared from Xcc A w -positive leaf samples. Xcc A is the DNA fraction prepared from a Xcc A positive sample. NC (negative control) was the healthy leaf DNA fraction. The ‘water’ sample indicates the LAMP reaction conducted with nuclease-free water. The control and test lines are indicated on the left, which detected the gold conjugate and Biotin- and DIG-labeled LAMP-A w products, respectively.

Article Snippet: In order to examine the field applicability of the LAMP-A w assay, the LAMP-A w reaction was conducted with crude DNA extracts prepared with 0.8% NaOH or AMP1 buffer (Agdia) following the manufacturer’s instructions, where the sample boiling step is not required for the crude DNA extract preparation.

Techniques: Negative Control, Control, Labeling