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Evaluation of filter paper to transport inactivated bacteria to detect carbapenem resistance genes by multiplex real-time PCR using high-resolution melting
- Source :
- Braz J Microbiol
- Publication Year :
- 2021
- Publisher :
- Springer Science and Business Media LLC, 2021.
-
Abstract
- Infections caused by resistant microorganisms are a complex global public health challenge, and the way to combat the increase of resistance is the development of more modern and faster techniques for resistance detection. This study aimed to evaluate the transport of inactivated bacteria impregnated in a filter paper disk to detect carbapenem resistance genes by multiplex real-time PCR (qPCR) using high-resolution melting (HRM). A total of 88 isolates of 10 different species of Enterobacterales harboring well-characterized carbapenem resistance genes were evaluated. A full 10-µL loop of fresh growth of bacteria were impregnated in a filter paper disk, which was left at room temperature for 2 days in order to simulate the time spent in transportation. Bacterial inactivation was performed with 70% ethanol at 15 min. Afterwards, the DNA was extracted from the paper disks for further analysis by qPCR HRM. The time of 15 min in 70% ethanol was enough to inactivate all the isolates tested. It was possible to correctly identify the presence of the carbapenem resistance gene by HRM qPCR in 87 isolates (98.87%) that were transported in the filter paper disks. Our results indicated that it is possible to use filter paper to transport inactivated bacteria and to identify carbapenem resistance genes by qPCR HRM. This alternative tends to facilitate the access to this technology by many laboratories which do not have the qPCR equipment.
- Subjects :
- Paper
medicine.medical_specialty
Bacteria
Ethanol
Filter paper
Microorganism
Biology
Real-Time Polymerase Chain Reaction
biology.organism_classification
Microbiology
High Resolution Melt
Specimen Handling
Real-time polymerase chain reaction
Medical microbiology
Carbapenems
Drug Resistance, Bacterial
Media Technology
medicine
Food microbiology
Multiplex
Clinical Microbiology - Short Communication
Subjects
Details
- ISSN :
- 16784405 and 15178382
- Volume :
- 52
- Database :
- OpenAIRE
- Journal :
- Brazilian Journal of Microbiology
- Accession number :
- edsair.doi.dedup.....be6ac7e1f20abb97466ddd1b5f59f7de