Antibiotics, Vol. 14, Pages 772: Finding the Missing IMP Gene: Overcoming the Imipenemase IMP Gene Drop-Out in Automated Molecular Testing for Carbapenem-Resistant Bacteria Circulating in Latin America

Antibiotics doi: 10.3390/antibiotics14080772

Authors:
Jose Arturo Molina-Mora
Ángel Rojas-Varela
Christopher Martínez-Arana
Lucia Portilla-Victor
Isaac Quirós-Fallas
Miryana Sánchez-Fonseca
Xavier Araya
Daniel Cascante-Serrano
Elvira Segura-Retana
Carlos Espinoza-Solís
María Jose Uribe-Calvo
Vanessa Villalobos-Alfaro
Heylin Estrada-Murillo
Stephanie Montoya-Madriz
Warren Madrigal
Mauricio Lizano
Stefany Lozada-Alvarado
Mariela Alvarado-Rodríguez
Mauricio Bolaños-Muñoz
Cristina García-Marín
Javier Alfaro-Camacho
Gian Carlo González-Carballo
Leana Quirós-Rojas
Joseph Sánchez-Fernández
Carolina Chaves-Ulate
Fernando García-Santamaría

Carbapenem resistance is considered one of the greatest current threats to public health, particularly in the management of infections in clinical settings. Carbapenem resistance in bacteria is mainly due to mechanisms such as the production of carbapenemases (such as the imipenemase IMP, or other enzymes like VIM, NDM, and KPC), that can be detected by several laboratory tests, including immunochromatography and automated real-time PCR (qPCR). Methods: As part of local studies to monitor carbapenem-resistant bacteria in Costa Rica, two cases were initially identified with inconsistent IMP detection results. A possible gene drop-out in the automated qPCR test was suggested based on the negative result, contrasting with the positive result by immunochromatography and whole-genome sequencing. We hypothesized that molecular testing could be optimized through the development of tailored assays to improve the detection of IMP genes. Thus, using IMP gene sequences from the local isolates and regional sequences in databases, primers were redesigned to extend the detection of IMP alleles of regional relevance. Results: The tailored qPCR was applied to a local collection of 119 carbapenem-resistant isolates. The genomes of all 14 positive cases were sequenced, verifying the results of the custom qPCR, despite the negative results of the automated testing. Conclusions: Guided by whole-genome sequencing, it was possible to extend the molecular detection of IMP alleles circulating in Latin America using a tailored qPCR to overcome IMP gene drop-out and false-negative results in an automated qPCR.

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Jose Arturo Molina-Mora www.mdpi.com