Journal: Microorganisms

Article Title: On-Site Dual Detection of Airborne Acinetobacter baumannii and Its Carbapenem-Resistant Gene bla OXA-23 Using a One-Pot Visual LAMP-CRISPR/Cas12a-Based Platform

doi: 10.3390/microorganisms13050976

Figure Lengend Snippet: Optimization of the one-pot LAMP-CRISPR/Cas12a assay targeting A. baumannii . ( a ) Fluorescence changed over time in the one-pot LAMP-CRISPR/Cas12a assay with a 1:1 ratio of Cas12a and crRNA. ( b ) Fluorescence signals in the one-pot LAMP-CRISPR/Cas12a assays with varying concentrations of crRNA. ( c ) Fluorescence signals in the one-pot LAMP-CRISPR/Cas12a assays with varying concentrations of FAM-labeled reporter probe. All concentrations in this figure referred to final concentration in the total 50 μL LAMP-CRISPR/Cas12a assay, comprising a 25 μL LAMP reaction and a 25 μL CRISPR/Cas12a reaction system, excluding 20 μL of mineral oil. The error bars represent mean + standard deviation ( n = 3 replicates). Bars labeled with different letters indicate statistically significant differences ( p < 0.05). Shared letters (e.g., ‘a, ab’) denote no significant difference between groups.

Article Snippet: The CRISPR/Cas12a reaction mixture contained RNase-free water, 2× NEB buffer r2.1, 50 U RRI, 50 nM to 200 nM crRNA, 60 nM to 300 nM Lba Cas12a, 200 nM to 2 μM FAM- or ROX-labeled reporter probes.

Techniques: CRISPR, Fluorescence, Labeling, Concentration Assay, Standard Deviation

Journal: Microorganisms

Article Title: On-Site Dual Detection of Airborne Acinetobacter baumannii and Its Carbapenem-Resistant Gene bla OXA-23 Using a One-Pot Visual LAMP-CRISPR/Cas12a-Based Platform

doi: 10.3390/microorganisms13050976

Figure Lengend Snippet: Feasibility and specificity of the one-pot LAMP-CRISPR/Cas12a assay. ( a ) Target: A. baumannii . 1, A. baumannii (ATCC 19606); 2, A. baumannii CI-1; 3, A. baumannii CI-2; 4, A. baumannii CI-3; 5, A. baumannii CI-4; 6, A. baumannii CI-5; 7, A. baumannii CI-6; 8, A. baumannii CI-7; 9, A. baumannii CI-8; 10, A. baumannii EI-1; 11, A. baumannii EI-2; 12, Staphylococcus aureus ; 13, Staphylococcus epidermidis ; 14, Streptococcus pneumoniae ; 15, Escherichia coli ; 16, Pseudomonas aeruginosa ; 17, Klebsiella pneumoniae ; 18, Candida albicans ; 19, Aspergillus fumigatus ; 20, Cryptococcus neoformans . ( b ) Target: bla OXA-23 gene. 1, A. baumannii CI-1; 2, A. baumannii CI-2; 3, A. baumannii CI-3; 4, A. baumannii CI-4; 5, A. baumannii CI-5; 6, A. baumannii CI-6; 7, A. baumannii CI-7; 8, A. baumannii CI-8; 9, A. baumannii (ATCC 19606); 10, CSAB 1; 11, CSAB 2; 12, CSAB 3. CI: clinical isolate; EI: environmental isolate; CSAB: Carbapenem-Susceptible A. baumannii ; A. baumannii CI 1-8 were A. baumannii isolates that carried the bla OXA-23 gene. CSAB 1-3 were confirmed to be devoid of the bla OXA-23 gene.

Article Snippet: The CRISPR/Cas12a reaction mixture contained RNase-free water, 2× NEB buffer r2.1, 50 U RRI, 50 nM to 200 nM crRNA, 60 nM to 300 nM Lba Cas12a, 200 nM to 2 μM FAM- or ROX-labeled reporter probes.

Techniques: CRISPR

Journal: Microorganisms

Article Title: On-Site Dual Detection of Airborne Acinetobacter baumannii and Its Carbapenem-Resistant Gene bla OXA-23 Using a One-Pot Visual LAMP-CRISPR/Cas12a-Based Platform

doi: 10.3390/microorganisms13050976

Figure Lengend Snippet: Sensitivity of the one-pot LAMP-CRISPR/Cas12a assay. ( a , b ) Relative fluorescence changed within 5 min. ( c , d ) Relative fluorescence values after 5 min of Cas12a-mediated cleavage. Error bars showed mean + standard deviation ( n = 3 replicates). ( e , f ) Direct observation by the naked eye under UV light (312 nm). ( a , c , e ) Target: A. baumannii . ( b , d , f ) Target: bla OXA-23 . NC, negative control.

Article Snippet: The CRISPR/Cas12a reaction mixture contained RNase-free water, 2× NEB buffer r2.1, 50 U RRI, 50 nM to 200 nM crRNA, 60 nM to 300 nM Lba Cas12a, 200 nM to 2 μM FAM- or ROX-labeled reporter probes.

Techniques: CRISPR, Fluorescence, Standard Deviation, Negative Control