Single-Tube Multiplex Polymerase Chain Reaction for the Detection of Genes Encoding Enterobacteriaceae Carbapenemase

CRISPR-Cas13a-Based Assay for Accurate Detection of OXA-48 and GES Carbapenemases

“…Comparison of the sensitivity and specificity of our assay with those obtained with other assays showed that our results are at least as good as those of other LAMP assays and most multiplex qPCR assays ( 18 , 21 – 24 , 26 – 28 ). In addition, we obtained higher specificity values than the Check-Direct CPE ( 25 ) and Luminex xTAG assay ( 19 ), as well as higher sensitivity values than the Amplidiag CarbaR+MCR kit ( 20 ) (Table S1).…”

“…A total of 12 studies on molecular diagnostics of different carbapenemases were found; the bla OXA-48 gene was detected in all of these studies, and the bla GES gene was detected in seven of them ( 18 – 24 ). Regarding the diagnostic technologies on which these techniques were based, seven out of 12 articles were based on multiplex qPCR (Check Direct CPE, Amplidiag CarbaR+MCR Kit, EntericBio CPE, BD Max System and in-house reactions) ( 20 – 26 ), two out of 12 protocols were based on LAMP reaction (Eazyplex SuperBug CRE and LAMP-HNB) ( 27 , 28 ), and three out of 12 were based on Luminex system (Luminex xTAG) ( 19 ), multiplex conventional PCR (in-house reaction) ( 18 ) and multiplex microarray technology, respectively ( 29 ).…”

“…In addition, five out of seven multiplex qPCR assays ( 21 – 24 , 26 ) yielded excellent results, with parameter values of 100%, although they were slower (1 to 3 h). Likewise, very promising results were achieved with Luminex technology and even multiplex conventional PCR (specificity of 99.4% to 100% and sensitivity of 100%), although the assays were also slower (2 to 5 h) ( 18 , 19 ). By contrast, poorer results were obtained with the other two multiplex qPCR assays (Amplidiag CarbaR+MCR and Check-Direct CPE) ( 20 , 25 ), which yielded sensitivity of 92% to 100% and specificity of 88 to 100, and took up to 3 h. Finally, multiplex microarray-based technology yielded a sensitivity of 98.3%, specificity of 99.6%; and was faster than multiplex qPCR (2 h) ( 29 ).…”

Carbapenemase-producing Enterobacterales isolated from hospital sinks: molecular relationships with isolates from patients and the change in contamination status after daily disinfection with sodium hypochlorite

“…To detect carbapenemase, we used Cica-βtest MBL and MASTDISCS combi Carba plus disc system (MD). 7 Carbapenemase genes were identified using multiplex polymerase chain reaction, according to the methods described by Watahiki et al 8 Molecular characterization of the outbreak and environmental isolates was performed using pulse-field gel electrophoresis (PFGE). Genomic DNA of Pantoea sp., Enterobacter cloacae complex, and Klebsiella oxytoca were processed using Xba I, Spe I, and Xba I, respectively.…”