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Microorganisms
Volume 9
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10.3390/microorganisms9051031
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Article

Validation of a Novel Diagnostic Approach Combining the VersaTREK™ System for Recovery and Real-Time PCR for the Identification of Mycobacterium chimaera in Water Samples

by
Roberto Zoccola
1,
Alessia Di Blasio
1,2,*,
Tiziana Bossotto
1,
Angela Pontei
1,
Maria Angelillo
1,
Alessandro Dondo
1,
Maria Goria
1 and
Simona Zoppi
1
1
Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, 10154 Torino, Italy
2
Azienda Sanitaria Locale TO3 S.C. Sanità Animale, Pinerolo, 10064 Torino, Italy
*
Author to whom correspondence should be addressed.
Microorganisms 2021, 9(5), 1031; https://doi.org/10.3390/microorganisms9051031
Submission received: 7 April 2021 / Revised: 3 May 2021 / Accepted: 9 May 2021 / Published: 11 May 2021
(This article belongs to the Special Issue Nontuberculous Mycobacteria: Ecology and Impact on Animal and Human Health)
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Abstract

Mycobacterium chimaera is an emerging pathogen associated with endocarditis and vasculitis following cardiac surgery. Although it can take up to 6–8 weeks to culture on selective solid media, culture-based detection remains the gold standard for diagnosis, so more rapid methods are urgently needed. For the present study, we processed environmental M. chimaera infected simulates at volumes defined in international guidelines. Each preparation underwent real-time PCR; inoculates were placed in a VersaTREK™ automated microbial detection system and onto selective Middlebrook 7H11 agar plates. The validation tests showed that real-time PCR detected DNA up to a concentration of 10 ng/µL. A comparison of the isolation tests showed that the PCR method detected DNA in a dilution of ×102 CFU/mL in the bacterial suspensions, whereas the limit of detection in the VersaTREK™ was <10 CFU/mL. Within less than 3 days, the VersaTREK™ detected an initial bacterial load of 100 CFU. The detection limit did not seem to be influenced by NaOH decontamination or the initial water sample volume; analytical sensitivity was 1.5 × 102 CFU/mL; positivity was determined in under 15 days. VersaTREK™ can expedite mycobacterial growth in a culture. When combined with PCR, it can increase the overall recovery of mycobacteria in environmental samples, making it potentially applicable for microbial control in the hospital setting and also in environments with low levels of contamination by viable mycobacteria.
Keywords: non-tuberculous mycobacteria; culture media; limit of detection; real-time polymerase chain reaction non-tuberculous mycobacteria; culture media; limit of detection; real-time polymerase chain reaction

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MDPI and ACS Style

Zoccola, R.; Di Blasio, A.; Bossotto, T.; Pontei, A.; Angelillo, M.; Dondo, A.; Goria, M.; Zoppi, S. Validation of a Novel Diagnostic Approach Combining the VersaTREK™ System for Recovery and Real-Time PCR for the Identification of Mycobacterium chimaera in Water Samples. Microorganisms 2021, 9, 1031. https://doi.org/10.3390/microorganisms9051031

AMA Style

Zoccola R, Di Blasio A, Bossotto T, Pontei A, Angelillo M, Dondo A, Goria M, Zoppi S. Validation of a Novel Diagnostic Approach Combining the VersaTREK™ System for Recovery and Real-Time PCR for the Identification of Mycobacterium chimaera in Water Samples. Microorganisms. 2021; 9(5):1031. https://doi.org/10.3390/microorganisms9051031

Chicago/Turabian Style

Zoccola, Roberto, Alessia Di Blasio, Tiziana Bossotto, Angela Pontei, Maria Angelillo, Alessandro Dondo, Maria Goria, and Simona Zoppi. 2021. "Validation of a Novel Diagnostic Approach Combining the VersaTREK™ System for Recovery and Real-Time PCR for the Identification of Mycobacterium chimaera in Water Samples" Microorganisms 9, no. 5: 1031. https://doi.org/10.3390/microorganisms9051031

APA Style

Zoccola, R., Di Blasio, A., Bossotto, T., Pontei, A., Angelillo, M., Dondo, A., Goria, M., & Zoppi, S. (2021). Validation of a Novel Diagnostic Approach Combining the VersaTREK™ System for Recovery and Real-Time PCR for the Identification of Mycobacterium chimaera in Water Samples. Microorganisms, 9(5), 1031. https://doi.org/10.3390/microorganisms9051031

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