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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

  • Clinical Microbiology - Short Communication
  • Published:
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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.

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Funding

The authors received funding from INPRA—Instituto Nacional de Pesquisa em Resistência Antimicrobiana—Brazil (INCT/CNPq: 465718/2014–0 and INCT/FAPERGS: 17/2551–0000514-7) and from Fundo de Incentivo a Pesquisa e Eventos do Hospital de Clínicas de Porto Alegre (FIPE/HCPA).

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Authors and Affiliations

  1. LABRESIS - Laboratório de Pesquisa Em Resistência Bacteriana, Hospital de Clínicas de Porto Alegre 2350, Porto Alegre, Rio Grande Do Sul, 90.035-903, Brazil

    M. S. Carneiro, M. N. Crispim, Priscila Lamb Wink & A. L. Barth

  2. PPGCF - Programa de Pós-Graduação Em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal Do Rio Grande Do Sul, Porto Alegre, Rio Grande do Sul, Brazil

    M. S. Carneiro, Priscila Lamb Wink & A. L. Barth

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  1. M. S. Carneiro
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  2. M. N. Crispim
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  3. Priscila Lamb Wink
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  4. A. L. Barth
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Contributions

Conceptualization: Maiara dos Santos Carneiro; methodology: Maiara dos Santos Carneiro; formal analysis and investigation: Maiara dos Santos Carneiro, Marina Niada Crispim, and Priscila Lamb Wink; writing—original draft preparation: Maiara dos Santos Carneiro; writing—review and editing: Maiara dos Santos Carneiro, Priscila Lamb Wink, and Afonso Luis Barth; funding acquisition: Afonso Luis Barth; resources: Afonso Luis Barth; and supervision: Afonso Luis Barth.

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Correspondence to Priscila Lamb Wink.

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Approval was obtained from the ethics committee of Hospital de Clínicas de Porto Alegre (CAAE: 41738520.0.0000.5327).

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The authors declare no competing interests.

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Carneiro, M.S., Crispim, M.N., Wink, P.L. et al. Evaluation of filter paper to transport inactivated bacteria to detect carbapenem resistance genes by multiplex real-time PCR using high-resolution melting . Braz J Microbiol 52, 1353–1356 (2021). https://doi.org/10.1007/s42770-021-00530-2

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  • DOI: https://doi.org/10.1007/s42770-021-00530-2

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