Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid and Specific Identification of Carbapenem-Resistant Acinetobacter baumannii Strains Harboring blaOXA-23, and the Epidemiological Survey of Clinical Isolates

2020 ◽  
Vol 26 (12) ◽  
pp. 1458-1465
Author(s):  
Puyuan Li ◽  
Wenkai Niu ◽  
Yun Fang ◽  
Dayang Zou ◽  
Huiying Liu ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Epidemiological Survey ◽  
Isothermal Amplification ◽  
Clinical Isolates ◽  
Specific Identification ◽  
Loop Mediated Isothermal Amplification ◽  
Carbapenem Resistant ◽  
Amplification Assay

scholarly journals Evaluation of a Loop-Mediated Isothermal Amplification assay to detect carbapenemases from Acinetobacter spp. directly from bronchoalveolar lavage fluid

2020 ◽  
Author(s):  
J. Moreno-Morales ◽  
A. Vergara ◽  
T. Kostyanev ◽  
J. Rodriguez-Baño ◽  
H. Goossens ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Acinetobacter Baumannii ◽  
Nosocomial Infections ◽  
Bronchoalveolar Lavage Fluid ◽  
Lavage Fluid ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Carbapenem Resistant ◽  
Amplification Assay ◽  
Acinetobacter Spp

AbstractCarbapenem-resistant Acinetobacter spp. mainly Acinetobacter baumannii are frequently causing nosocomial infections with high mortality. In this study, the efficacy of the Eazyplex® SuperBug Complete A system (Amplex Diagnostics GmbH, Gars-Bahnhof, Germany), based on loop-mediated isothermal amplification (LAMP), to detect the presence of carbapenemases in Acinetobacter spp. directly from bronchoalveolar lavage samples was assessed, detecting all tested carbapenemases in less than 30 minutes with a sensitivity of 103 CFU/ml.

scholarly journals Evaluation of a Loop-Mediated Isothermal Amplification-Based Methodology To Detect Carbapenemase Carriage in Acinetobacter Clinical Isolates

2014 ◽  
Vol 58 (12) ◽  
pp. 7538-7540 ◽  
Author(s):  
Andrea Vergara ◽  
Yuliya Zboromyrska ◽  
Noraida Mosqueda ◽  
María Isabel Morosini ◽  
Sergio García-Fernández ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Nosocomial Infections ◽  
Isothermal Amplification ◽  
Clinical Isolates ◽  
Content Type ◽  
Loop Mediated Isothermal Amplification ◽  
System A ◽  
Carbapenem Resistant ◽  
Different Types ◽  
Hydrolyzing Enzymes

ABSTRACTCarbapenem-resistantAcinetobacter baumanniiis a major source of nosocomial infections worldwide and is mainly associated with the acquisition of OXA-type carbapenemases and, to a lesser extent, metallo-β-lactamases (MBLs). In this study, 82 nonepidemiologically relatedAcinetobacterstrains carrying different types of OXA or MBL enzymes were tested using the Eazyplex system, a loop-mediated isothermal amplification (LAMP)-based method to rapidly detect carbapenemase carriage. The presence/absence of carbapenem-hydrolyzing enzymes was correctly determined for all isolates in <30 min.

scholarly journals Antibacterial effect of acetic acid during an outbreak of carbapenem-resistant Acinetobacter baumannii in an ICU (II)

2021 ◽  
Vol 15 (08) ◽  
pp. 1167-1172
Author(s):  
Carolina Garciglia-Mercado ◽  
Ramon Gaxiola-Robles ◽  
Felipe Ascencio ◽  
Concepción Grajales-Muñiz ◽  
Maria Luisa Soriano Rodríguez ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Acinetobacter Baumannii ◽  
Environmental Samples ◽  
Antibacterial Effect ◽  
Companion Paper ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Carbapenem Resistant ◽  
Before And After ◽  
Amplification Assay

Introduction: Acetic acid (AA) has been commonly used in medicine as an antiseptic agent for the past 6000 years. This study evaluated the antibacterial effect of AA during an outbreak in an intensive care unit (ICU) facility in Baja California Sur, México. Methodology: Thirty-five environmental samples were collected, subsequently, disinfection with AA (4%) was performed, and two days later the same areas were sampled inside the ICU facility. Carbapenem-resistant A. baumannii (CRAB) was detected with loop-mediated isothermal amplification assay (Garciglia-Mercado et al. companion paper), targeting blaOXA-23-like, blaOXA-24-like, blaOXA-51-like, blaOXA-58-like, blaIMP and blaVIM genes. CRAB isolates before and after disinfection were compared by PFGE. Results: Eighteen (54.5%) and five (14.3%) of thirty-five environmental samples were identified as Acinetobacter baumannii before and after disinfection, respectively, showing a significant decrease of 85.7% (p < 0.05) both by Loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR). Furthermore, the presence of blaOXA-23-like and blaOXA-58-like genes significantly decreased (p < 0.05) both by LAMP and PCR methods. PFGE genotype showed high similarity among CRAB isolates before and after disinfection, suggesting wide clonal dissemination in the ICU facility. Conclusions: This study demonstrated the novel application of AA with the LAMP assays developed for detecting CRAB. AA promises to be a cheap and efficacious disinfectant alternative to both developed and especially developing countries, preventing the spread of this organism in the environment and to other susceptible patients in health care settings.

scholarly journals Development of Loop-Mediated Isothermal Amplification Assay for Detection of Clinically Significant Members of Acinetobacter calcoaceticus–baumannii Complex and Associated Carbapenem Resistance

2021 ◽  
Vol 8 ◽  
Author(s):  
Amit Sharma ◽  
Rajni Gaind
Keyword(s):  
Acinetobacter Baumannii ◽  
Lamp Assay ◽  
Carbapenem Resistance ◽  
Acinetobacter Calcoaceticus ◽  
Isothermal Amplification ◽  
Colony Count ◽  
Dna Concentration ◽  
Loop Mediated Isothermal Amplification ◽  
Carbapenem Resistant ◽  
Clinically Significant

Background:Acinetobacter calcoaceticus–baumannii (ACB) complex has emerged as an important nosocomial pathogen and is associated with life-threatening infections, especially among ICU patients, including neonates. Carbapenem resistance in Acinetobacter baumannii has emerged globally and is commonly mediated by blaOXA-23. Clinically significant infections with carbapenem-resistant Acinetobacter baumannii (CRAB) are a major concern since therapeutic options are limited and associated mortality is high. Early diagnosis of both the pathogen and resistance is important to initiate the optimal therapy and prevent selection of resistance. In the current study, a loop-mediated isothermal amplification (LAMP) assay was developed for rapid detection of the ACB complex and carbapenem resistance mediated by blaOXA-23.Methodology: Universal LAMP primers were designed for the detection of significant members of the ACB complex and carbapenem resistance targeting the ITS 16S–23S rRNA and blaOXA-23 gene respectively. The optimal conditions for the LAMP assay were standardized for each primer set using standard ATCC strains. The sensitivity of the LAMP assay was assessed based on the limit of detection (LOD) using different DNA concentrations and colony counts. The specificity of LAMP was determined using the non-ACB complex and non-Acinetobacter species. The results of the LAMP assay were compared with those of polymerase chain reaction (PCR).Results: The optimal temperature for the LAMP assay was 65°C, and the detection time varied with various primers designed. Using the ITS Ab1 primer, LODs of LAMP and PCR assays were 100 pg/μl and 1 ng/μl of DNA concentration and 104 cfu/ml and 108 cfu/ml of colony count, respectively. The LAMP assay was 10- and 104-fold more sensitive than PCR using DNA concentration and colony count, respectively. The LAMP assay was found to be specific for clinically important ACB complex species.Significance of the study: The LAMP assay can be applied for early detection of significant species of the ACB complex from clinical samples and their carbapenem-resistant variants. Depending on the emerging pathogen and locally prevalent resistance genes, the LAMP assay can be modified for detection of colonization or infection by various resistant bugs.

scholarly journals Evaluation of a Loop-Mediated Isothermal Amplification Assay to Detect Carbapenemases Directly From Bronchoalveolar Lavage Fluid Spiked With Acinetobacter spp.

2021 ◽  
Vol 11 ◽  
Author(s):  
Javier Moreno-Morales ◽  
Andrea Vergara ◽  
Tomislav Kostyanev ◽  
Jesús Rodriguez-Baño ◽  
Herman Goossens ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Molecular Diagnostics ◽  
Bronchoalveolar Lavage Fluid ◽  
Lavage Fluid ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Carbapenem Resistant ◽  
Amplification Assay ◽  
Acinetobacter Spp ◽  
Selection Of

Carbapenem-resistant Acinetobacter spp. mainly Acinetobacter baumannii are frequently causing nosocomial infections with high mortality. In this study, the efficacy of the Eazyplex® SuperBug Complete A system, based on loop-mediated isothermal amplification (LAMP), to detect the presence of carbapenemases in Acinetobacter spp. directly from bronchoalveolar lavage (BAL) samples was assessed. A total of 22 Acinetobacter spp. strains producing OXA-23, OXA-40, OXA-58, NDM, and IMP were selected. Eazyplex SuperBug Complete A kit, used with the Genie II device, is a molecular diagnostics kit that detects a selection of genes that express carbapenemases (blaKPC, blaNDM, blaVIM, blaOXA–48, blaOXA–23, blaOXA–40, and blaOXA–58). Negative BAL samples were identified, McFarland solutions were prepared from each of the 22 Acinetobacter strains and serial dilutions in saline solution were made to finally spike BAL samples to a concentration of 102 and 103 CFU/ml. Fifteen concentrations out of the 44 tested out did not provide detection of the carbapenemase-producing gene, all but one being at the lowest concentration tested at 102 CFU/ml; therefore, the limit of sensitivity is 103 CFU/ml. This assay represents the kind of advantages that investing in molecular diagnostics brings to the clinical practice, allowing the identification of carbapenemases in less than 30 min with a sensitivity of 103 CFU/ml.

Performance of a loop-mediated isothermal amplification assay (Isoplex CRE-ART) to detect common carbapenemase-encoding genes in Gram-negative bacteria

2021 ◽  
Vol 70 (7) ◽  
Author(s):  
Laura Berneking ◽  
Lucia Asar ◽  
Anna Both ◽  
Benjamin Berinson ◽  
Martin Aepfelbacher ◽  
...  
Keyword(s):  
Isothermal Amplification ◽  
Gram Negative Bacteria ◽  
Pcr Assay ◽  
Gram Negative ◽  
Content Type ◽  
Loop Mediated Isothermal Amplification ◽  
Carbapenem Resistant ◽  
Amplification Assay ◽  
Pathogen Characterization ◽  
Encoding Genes

Carbapenem-resistant Gram-negative bacteria (CR-GNB) are a major source of nosocomial infections worldwide. In this study, the ability of a loop-mediated isothermal amplification (LAMP)-based method (Isoplex CRE-ART) to rapidly detect carbapenemase-encoding genes bla OXA-48-like, bla OXA-23-like, bla OXA-24-like, bla KPC, bla VIM, bla NDM and bla IMP in 231 carbapenem-resistant Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumannii isolates was investigated. The accuracy of the LAMP test was compared to results of molecular isolate characterization using a Laboratory Developed Test multiplex carbapenemase PCR assay. The LAMP test correctly identified the presence of on-panel carbapenemases with a sensitivity of 99.16 % [95 % confidence interval (CI): 95.39–99.96 %] and a specificity of 98.21 % (95 % CI: 93.72–99.68 %) in 60 min. Our findings suggest that the Isoplex CRE-ART assay is able to rapidly identify carbapenemase genes in CR-GNB and improves options for pathogen characterization in the context of clinical microbiological and infection control diagnostics.

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