1887

Abstract

. is an emerging fungal pathogen. The organism can cause invasive infections associated with high mortality, has been implicated in outbreaks in healthcare settings and is frequently resistant to multiple antifungal agents, making it a significant challenge to infection prevention and patient treatment.

To implement a real-time PCR assay for detection of in patient surveillance samples collected with the Copan Liquid Amies elution swab (ESwab) collection and transport system.

We optimized a real-time PCR testing procedure based on the sample collection device used in our institution.

ESwab transport medium was strongly inhibitory to the real-time PCR. Removing the medium with centrifugation, followed by suspending the pellet in PBS-BSA buffer (concentration 1 %), sufficiently eliminated the inhibition. The manual sample preparation method, freeze–thaw followed by mechanical disruption, allowed the detection of at the lowest cell concentration.

The optimized procedure was used to test 1414 patient surveillance samples. The real-time PCR detected all culture-positive samples with 100 % sensitivity and 100 % specificity.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001207
2020-06-01
2021-10-18
Loading full text...

Full text loading...

/deliver/fulltext/jmm/69/6/824.html?itemId=/content/journal/jmm/10.1099/jmm.0.001207&mimeType=html&fmt=ahah

References

  1. Jeffery-Smith A, Taori SK, Schelenz S, Jeffery K, Johnson EM et al. Candida auris: a review of the literature. Clin Microbiol Rev 2018; 31:e00029-17 [View Article][PubMed]
    [Google Scholar]
  2. CDC Clinical Alert to U.S. Healthcare Facilities – June 2016. Global Emergence of Invasive Infections Caused by the Multidrug-Resistant Yeast Candida auris Atlanta, GA: Centers for Disease Control and Prevention; 2016
    [Google Scholar]
  3. Schelenz S, Hagen F, Rhodes JL, Abdolrasouli A, Chowdhary A et al. First hospital outbreak of the globally emerging Candida auris in a European hospital. Antimicrob Resist Infect Control 2016; 5:35 [View Article][PubMed]
    [Google Scholar]
  4. Shackleton JSS, Rochon M, Hall A, Ryan L, Cervera-Jackson R. Impact of environmental decontamination in a Candida auris outbreak. J Hosp Infect 2016; 94:S24
    [Google Scholar]
  5. Vallabhaneni S, Kallen A, Tsay S, Chow N, Welsh R et al. Investigation of the first seven reported cases of Candida auris, a globally emerging invasive, multidrug-resistant fungus – United States, May 2013–August 2016. Am J Transplant 2017; 17:296–299 [View Article][PubMed]
    [Google Scholar]
  6. Wickes BL. Analysis of a Candida auris outbreak provides new insights into an emerging pathogen. J Clin Microbiol 2020; 58:e02083-19 [View Article][PubMed]
    [Google Scholar]
  7. Leach L, Zhu Y, Chaturvedi S. Development and validation of a real-time PCR assay for rapid detection of Candida auris from surveillance samples. J Clin Microbiol 2018; 56:e01223-17 [View Article][PubMed]
    [Google Scholar]
  8. Nummi M, Mannonen L, Puolakkainen M. Development of a multiplex real-time PCR assay for detection of Mycoplasma pneumoniae, Chlamydia pneumoniae and mutations associated with macrolide resistance in Mycoplasma pneumoniae from respiratory clinical specimens. Springerplus 2015; 4:684 [View Article][PubMed]
    [Google Scholar]
  9. White TJB, Lee T, Taylor S. PCR Protocols: a Guide to Methods and Applications London: Academic Press; 1990
    [Google Scholar]
  10. Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM et al. Rapid and simple method for purification of nucleic acids. J Clin Microbiol 1990; 28:495–503 [View Article][PubMed]
    [Google Scholar]
  11. Carbonero F, Nava GM, Benefiel AC, Greenberg E, Gaskins HR. Microbial DNA extraction from intestinal biopsies is improved by avoiding mechanical cell disruption. J Microbiol Methods 2011; 87:125–127 [View Article][PubMed]
    [Google Scholar]
  12. Salonen A, Nikkilä J, Jalanka-Tuovinen J, Immonen O, Rajilić-Stojanović M et al. Comparative analysis of fecal DNA extraction methods with phylogenetic microarray: effective recovery of bacterial and archaeal DNA using mechanical cell lysis. J Microbiol Methods 2010; 81:127–134 [View Article][PubMed]
    [Google Scholar]
  13. Murray AE, Preston CM, Massana R, Taylor LT, Blakis A et al. Seasonal and spatial variability of bacterial and archaeal assemblages in the coastal waters near Anvers Island, Antarctica. Appl Environ Microbiol 1998; 64:2585–2595 [View Article][PubMed]
    [Google Scholar]
  14. Fujita S, Lasker BA, Lott TJ, Reiss E, Morrison CJ. Microtitration plate enzyme immunoassay to detect PCR-amplified DNA from Candida species in blood. J Clin Microbiol 1995; 33:962–967 [View Article][PubMed]
    [Google Scholar]
  15. van Deventer AJ, Goessens WH, van Belkum A, van Vliet HJ, van Etten EW et al. Improved detection of Candida albicans by PCR in blood of neutropenic mice with systemic candidiasis. J Clin Microbiol 1995; 33:625–628 [View Article][PubMed]
    [Google Scholar]
  16. Karakousis A, Tan L, Ellis D, Alexiou H, Wormald PJ. An assessment of the efficiency of fungal DNA extraction methods for maximizing the detection of medically important fungi using PCR. J Microbiol Methods 2006; 65:38–48 [View Article][PubMed]
    [Google Scholar]
  17. Lech T, Syguła-Cholewinska J, Szostak-Kot J. An economical and combined method for rapid and efficient isolation of fungal DNA. Genet Mol Res 2014; 13:10779–10786 [View Article][PubMed]
    [Google Scholar]
  18. Shin JH, Nolte FS, Morrison CJ. Rapid identification of Candida species in blood cultures by a clinically useful PCR method. J Clin Microbiol 1997; 35:1454–1459 [View Article][PubMed]
    [Google Scholar]
  19. da Silva GA, Bernardi TL, Schaker PDC, Menegotto M, Valente P. Rapid yeast DNA extraction by boiling and freeze-thawing without using chemical reagents and DNA purification. Braz Arch Biol Technol 2012; 55:319–327 [View Article]
    [Google Scholar]
  20. Gibb AP, Wong S. Inhibition of PCR by agar from bacteriological transport media. J Clin Microbiol 1998; 36:275–276 [View Article][PubMed]
    [Google Scholar]
  21. Cendejas-Bueno E, Kolecka A, Alastruey-Izquierdo A, Theelen B, Groenewald M et al. Reclassification of the Candida haemulonii complex as Candida haemulonii (C. haemulonii group I), C. duobushaemulonii sp. nov. (C. haemulonii group II), and C. haemulonii var. vulnera var. nov.: three multiresistant human pathogenic yeasts. J Clin Microbiol 2012; 50:3641–3651 [View Article][PubMed]
    [Google Scholar]
  22. Kathuria S, Singh PK, Sharma C, Prakash A, Masih A et al. Multidrug-resistant Candida auris misidentified as Candida haemulonii: characterization by matrix-assisted laser desorption ionization-time of flight mass spectrometry and DNA sequencing and its antifungal susceptibility profile variability by Vitek 2, CLSI broth microdilution, and Etest method. J Clin Microbiol 2015; 53:1823–1830 [View Article][PubMed]
    [Google Scholar]
  23. Biswal M, Rudramurthy SM, Jain N, Shamanth AS, Sharma D et al. Controlling a possible outbreak of Candida auris infection: lessons learnt from multiple interventions. J Hosp Infect 2017; 97:363–370 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001207
Loading
/content/journal/jmm/10.1099/jmm.0.001207
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error