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Polymerase Chain Reaction Assay for Detection of Stenotrophomonas maltophilia in Cheese Samples Based on the smeT Gene

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Abstract

Stenotrophomonas maltophilia is an emerging opportunistic pathogen linked not only to bacteremia, sepsis, and pneumonia but also to severe chronic enteritis. Persons with the impaired immune system are prone to be infected by S. maltophilia since its pathogenicity seems to be more associated with the host immune system than with the acquisition of specific virulence genes. In the dairy chain, S. maltophilia is linked to clinical and subclinical bovine mastitis in dairy cows, and it has been identified in cheese, and raw and pasteurized milk. There are reports of misidentification of S. maltophilia by commercial systems and PCR assays using primers based on the 23S rRNA and smeD genes, so the smeT gene is an alternative to identifying S. maltophilia by PCR due to its specificity to the S. maltophilia species. The present study reports an alternative species-specific PCR assay based on the smeT gene designed to identify S. maltophilia in cheese samples. We performed in silico and in vitro analyses to check the specificity of the primer pair. In silico analysis showed specificity of the primer pair to the species level. In vitro analysis was performed by testing the primer pair against pools of bacteria grown from 33 fresh Minas cheese samples acquired in the city of Rio de Janeiro, Brazil, without unspecific amplification.

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References

  1. Alonso A, Martínez JL (2000) Cloning and characterization of SmeDEF, a novel multidrug efflux pump from Stenotrophomonas maltophilia. Antimicrob Agents Chemother 44:3079–3086. https://doi.org/10.1128/AAC.44.11.3079-3086.2000

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Apisarnthanarak A, Fraser VJ, Dunne WM et al (2003) Stenotrophomonas maltophilia intestinal colonization in hospitalized oncology patients with diarrhea. Clin Infect Dis 37:1131–1135. https://doi.org/10.1086/378297

    Article  PubMed  Google Scholar 

  3. Bhanderi BB, Jhala MK, Ahir VB et al (2014) Cultural and metagenomic based identification of a microbiome from subclinical mastitis in cows. Vet Arh 84:215–228

    Google Scholar 

  4. Brooke JS (2012) Stenotrophomonas maltophilia: an emerging global opportunistic pathogen. Clin Microbiol Rev 25:2–41. https://doi.org/10.1128/CMR.00019-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Coton M, Delbés-Paus C, Irlinger F et al (2012) Diversity and assessment of potential risk factors of Gram-negative isolates associated with French cheeses. Food Microbiol 29:88–98. https://doi.org/10.1016/j.fm.2011.08.020

    Article  CAS  PubMed  Google Scholar 

  6. Delbès C, Ali-Mandjee L, Montel MC (2007) Monitoring bacterial communities in raw milk and cheese by culture-dependent and -independent 16S rRNA gene-based analyses. Appl Environ Microbiol 73:1882–1891. https://doi.org/10.1128/AEM.01716-06

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Denton M, Kerr KG (1998) Microbiological and clinical aspects of infection associated with Stenotrophomonas maltophilia. Clin Microbiol Rev 11:57–80

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Falagas ME, Kastoris AC, Vouloumanou EK et al (2009) Attributable mortality of Stenotrophomonas maltophilia infections: a systematic review of the literature. Future Microbiol 4:1103–1109. https://doi.org/10.2217/fmb.09.84

    Article  PubMed  Google Scholar 

  9. Foster NF, Harnett GB, Riley TV, Chang BJ (2008) Cross-reaction of Stenotrophomonas and Xanthomonas species in a 23S rRNA gene-directed PCR for detection of S. maltophilia. J Clin Microbiol 46:4111–4113. https://doi.org/10.1128/JCM.01604-08

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Fredslund J, Schauser L, Madsen LH et al (2005) PriFi: using a multiple alignment of related sequences to find primers for amplification of homologs. Nucleic Acids Res 33:W516–W520. https://doi.org/10.1093/nar/gki425

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. García-León G, Hernández A, Hernando-Amado S et al (2014) A function of SmeDEF, the major quinolone resistance determinant of Stenotrophomonas maltophilia, is the colonization of plant roots. Appl Environ Microbiol 80:4559–4565. https://doi.org/10.1128/AEM.01058-14

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Hagi T, Sasaki K, Aso H, Nomura M (2013) Adhesive properties of predominant bacteria in raw cow’s milk to bovine mammary gland epithelial cells. Folia Microbiol (Praha) 58:515–522. https://doi.org/10.1007/s12223-013-0240-z

    Article  CAS  Google Scholar 

  13. Hellmig S, Ott S, Musfeldt M et al (2005) Life-threatening chronic enteritis due to colonization of the small bowel with Stenotrophomonas maltophilia. Gastroenterology 129:706–712. https://doi.org/10.1016/j.gastro.2005.01.011

    Article  PubMed  Google Scholar 

  14. Hernández A, Maté MJ, Sánchez-Díaz PC et al (2009) Structural and functional analysis of SmeT, the repressor of the Stenotrophomonas maltophilia multidrug efflux pump SmeDEF. J Biol Chem 284:14428–14438. https://doi.org/10.1074/jbc.M809221200

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Hernández A, Ruiz FM, Romero A, Martínez JL (2011) The binding of triclosan to SmeT, the repressor of the multidrug efflux pump SmeDEF, induces antibiotic resistance in Stenotrophomonas maltophilia. PLoS Pathog 7:e1002103. https://doi.org/10.1371/journal.ppat.1002103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Juffs HS (1973) Identification of Pseudomonas spp. isolated from milk produced in South Eastern Queensland. J Appl Bacteriol 36:559–585. https://doi.org/10.1111/j.1365-2672.1973.tb04145.x

    Article  Google Scholar 

  17. Larkin MA, Blackshields G, Brown NP et al (2007) Clustal W and clustal X version 2.0. Bioinformatics 23:2947–2948. https://doi.org/10.1093/bioinformatics/btm404

    Article  CAS  PubMed  Google Scholar 

  18. Lira F, Berg G, Martínez JL (2017) Double-face meets the bacterial world: the opportunistic pathogen Stenotrophomonas maltophilia. Front Microbiol 8:2190. https://doi.org/10.3389/fmicb.2017.02190

    Article  PubMed  PubMed Central  Google Scholar 

  19. Mallet A, Guéguen M, Kauffmann F et al (2012) Quantitative and qualitative microbial analysis of raw milk reveals substantial diversity influenced by herd management practices. Int Dairy J 27:13–21. https://doi.org/10.1016/j.idairyj.2012.07.009

    Article  Google Scholar 

  20. Ohnishi M, Sawada T, Marumo K et al (2012) Antimicrobial susceptibility and genetic relatedness of bovine Stenotrophomonas maltophilia isolates from a mastitis outbreak. Lett Appl Microbiol 54:572–576. https://doi.org/10.1111/j.1472-765X.2012.03246.x

    Article  CAS  PubMed  Google Scholar 

  21. Pinot C, Deredjian A, Nazaret S et al (2011) Identification of Stenotrophomonas maltophilia strains isolated from environmental and clinical samples: a rapid and efficient procedure. J Appl Microbiol 111:1185–1193. https://doi.org/10.1111/j.1365-2672.2011.05120.x

    Article  CAS  PubMed  Google Scholar 

  22. Renye JA, Somkuti GA, Vallejo-Cordoba B et al (2008) Characterization of the microflora isolated from Queso Fresco made from raw and pasteurized milk. J Food Saf 28:59–75. https://doi.org/10.1111/j.1745-4565.2007.00095.x

    Article  CAS  Google Scholar 

  23. Sanchez P, Alonso A, Martinez JL (2004) Regulatory regions of smeDEF in Stenotrophomonas maltophilia strains expressing different amounts of the multidrug efflux pump SmeDEF. Antimicrob Agents Chemother 48:2274–2276. https://doi.org/10.1128/AAC.48.6.2274-2276.2004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Todaro M, Francesca N, Reale S et al (2011) Effect of different salting technologies on the chemical and microbiological characteristics of PDO Pecorino Siciliano cheese. Eur Food Res Technol 233:931–940. https://doi.org/10.1007/s00217-011-1593-7

    Article  CAS  Google Scholar 

  25. Ye J, Coulouris G, Zaretskaya I et al (2012) Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics 13:134. https://doi.org/10.1186/1471-2105-13-134

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Zhang Z, Schwartz S, Wagner L, Miller W (2000) A greedy algorithm for aligning DNA sequences. J Comput Biol 7:203–214. https://doi.org/10.1089/10665270050081478

    Article  CAS  PubMed  Google Scholar 

  27. Zhang L, Li XZ, Poole K (2001) SmeDEF multidrug efflux pump contributes to intrinsic multidrug resistance in Stenotrophomonas maltophilia. Antimicrob Agents Chemother 45:3497–3503. https://doi.org/10.1128/AAC.45.12.3497-3503.2001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This work was supported by CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) (Grant Number 1440268)—MSc scholarship for N. Okuno.

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Authors and Affiliations

  1. Graduate Program in Food and Nutrition (PPGAN), Nutrition School, Federal University of the State of Rio de Janeiro (UNIRIO), Dr. Xavier Sigaud Street, 290, Urca, Rio de Janeiro, RJ, 22240-040, Brazil

    Newton T. Okuno & Cristiane R. Silva

  2. Nutrition School, Federal University of the State of Rio de Janeiro (UNIRIO), Dr. Xavier Sigaud Street, 290, Urca, Rio de Janeiro, RJ, 22240-040, Brazil

    Isabela R. Freire & Renata T. R. S. Segundo

  3. Department of Food Science (DCA), Nutrition School, Federal University of the State of Rio de Janeiro (UNIRIO), Dr. Xavier Sigaud Street, 290, Urca, Rio de Janeiro, RJ, 22240-040, Brazil

    Cristiane R. Silva & Victor A. Marin

Authors
  1. Newton T. Okuno
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  2. Isabela R. Freire
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  3. Renata T. R. S. Segundo
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  4. Cristiane R. Silva
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Correspondence to Newton T. Okuno.

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Okuno, N.T., Freire, I.R., Segundo, R.T.R.S. et al. Polymerase Chain Reaction Assay for Detection of Stenotrophomonas maltophilia in Cheese Samples Based on the smeT Gene. Curr Microbiol 75, 1555–1559 (2018). https://doi.org/10.1007/s00284-018-1559-0

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  • DOI: https://doi.org/10.1007/s00284-018-1559-0

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