Annals of Microbiology

, Volume 63, Issue 2, pp 683–689 | Cite as

Quick identification and quantification of Proteus mirabilis by polymerase chain reaction (PCR) assays

  • Weiwei Zhang
  • Zongliang Niu
  • Kun Yin
  • Ping Liu
  • Lingxin ChenEmail author
Original Article


Proteus mirabilis is an opportunistic pathogen that can cause urinary tract infection in human beings. The accurate and rapid identification and quantification of P. mirabilis is necessary for early treatment. In this study, a pair of specific primers according to the conserved ureR sequence of P. mirabilis was designed and novel systems which consisted of a polymerase chain reaction (PCR) and a real-time PCR to identify and quantify P. mirabilis were developed. For the qualitative identification by ordinary PCR, a 225-bp DNA product was amplified from P. mirabilis and separated on an agarose gel. The corresponding DNA product is present in three P. mirabilis strains isolated from different geographical locations, but is absent in 20 strains representing 18 different species, including the ureR homolog contained Providencia stuartii and Escherichia coli strains, the other common pathogens Klebsiella sp., Edwarsiella sp., Vibrio sp., Enterobacter sp., and Escherichia sp., and other environmental bacteria Pseudomonas sp. and Acinetobacter sp. Proteus mirabilis at concentrations higher than 1.0 × 103 CFU ml−1 was detectable by ordinary PCR; P. mirabilis at concentrations higher than 10 CFU ml−1 was quantified by real-time PCR. The specific, sensitive and time-efficient PCR methods were demonstrated to be applicable to rapid identification and quantification of P. mirabilis.


Proteus mirabilis ureR Polymerase chain reaction (PCR) Real-time PCR 



The authors are grateful for the financial support provided by the Innovation Projects of the Chinese Academy of Sciences grant KZCX2-EW-206, the National Natural Science Foundation of China (NSFC) grant 20975089, the Department of Science and Technology of Yantai City of China grant 2010235, the Doctoral Foundation of Shandong Province grant BS2011SW056, and the 100 Talents Program of the Chinese Academy of Sciences.


  1. Abolmaaty A, Gu W, Witkowsky R, Levin RE (2007) The use of activated charcoal for the removal of PCR inhibitors from oyster samples. J Microbiol Methods 68:349–352PubMedCrossRefGoogle Scholar
  2. Amri AA, Senok AC, Ismaeel AY, Al-Mahmeed AE, Botta GA (2007) Multiplex PCR for direct identification of Campylobacter spp. in human and chicken stools. J Med Microbiol 56:1350–1355PubMedCrossRefGoogle Scholar
  3. Anbazhagan D, Kathirvalu GG, Mansor M, Siok Yan GO, Yusof MY, Sekaran SD (2010) Multiplex polymerase chain reaction (PCR) assays for the detection of Enterobacteriaceae in clinical samples. Afi J Microbiol Res 4:1186–1191Google Scholar
  4. Brolund A, Wisell KT, Edquist PJ, Elfström L, Walder M, Giske CG (2010) Development of a real-time SYBRGreen PCR assay for rapid detection of acquired AmpC in Enterobacteriaceae. J Microbiol Meth 82:229–233CrossRefGoogle Scholar
  5. Cheng JC, Huang CL, Lin CC, Chen CC, Chang YC, Chang SS, Tseng CP (2006) Rapid detection and identification of clinically important bacteria by high-resolution melting analysis after broad-range ribosomal RNA real-time PCR. Clin Chem 52:1997–2004PubMedCrossRefGoogle Scholar
  6. Colwell RR (2009) Viable but not cultivable bacteria. Microbiol Monogr 10:121–129CrossRefGoogle Scholar
  7. D’Orazio SEF, Collins CM (1993) The plasmid-encoded urease gene cluster of the Enterobacteriaceae is positively regulated by UreR, a member of the AraC family of transcriptional activators. J Bacteriol 175:3459–3467PubMedGoogle Scholar
  8. dos Santos LR, do Nascimento VP, de Oliveira SD, Flores ML, Pontes AP, Ribeiro AR, Salle CT, Lopes RF (2001) Polymerase chain reaction (PCR) for the detection of Salmonella in artificially inoculated chicken meat. Rev Inst Med Trop Sao Paulo 43:247–250PubMedCrossRefGoogle Scholar
  9. Fiume L, Bucci Sabattini MA, Poda G (2005) Detection of Legionella pneumophila in water samples by species-specific real-time and nested PCR assays. Lett Appl Microbiol 41:470–475PubMedCrossRefGoogle Scholar
  10. Goarant C, Merien F (2006) Quantification of Vibrio penaeicida, the etiological agent of Syndrome 93 in New Caledonian shrimp, by real-time PCR using SYBR Green I chemistry. J Microbiol Meth 67:27–35CrossRefGoogle Scholar
  11. Hagi T, Kobayashi M, Nomura M (2010) Molecular-based analysis of changes in indigenous milk microflora during the grazing period. Biosci Biotechnol Biochem 74:484–487PubMedCrossRefGoogle Scholar
  12. Hooton TM (2003) Fluoroquinolones and resistance in the treatment of uncomplicated urinary tract infection. Int J Antimicrob Agents 22:65–72PubMedCrossRefGoogle Scholar
  13. Hryniewicz K, Szczypa K, Sulikowska A, Jankowski K, Betlejewska K, Hryniewicz W (2001) Antibiotic susceptibility of bacterial strains isolated from urinary tract infections in Poland. J Antimicrob Chemother 47:773–780PubMedCrossRefGoogle Scholar
  14. Huang HS, Chen J, Teng LJ, Lai MK (1999) Use of polymerase chain reaction to detect Proteus mirabilis and Ureaplasma urealyticum in urinary calculi. J Formos Med Assoc 98:844–850PubMedGoogle Scholar
  15. Le Dréan G, Mounier J, Vasseur V, Arzur D, Habrylo O, Barbier G (2010) Quantification of Penicillium camemberti and P. roqueforti mycelium by real-time PCR to assess their growth dynamics during ripening cheese. Int J Food Microbiol 138:100–107PubMedCrossRefGoogle Scholar
  16. Limanskiĭ A, Minukhin V, Limanskaia O, Pavlenko N, Mishina M, Tsygenenko A (2005) Species-specific detection of Proteus vulgaris and Proteus mirabilis by the polymerase chain reaction. Zh Mikrobiol Epidemiol Immunobiol 3:33–39PubMedGoogle Scholar
  17. Lu JJ, Perng CLH, Lee SY, Wan CC (2000) Use of PCR with universal primers and restriction endonuclease digestions for detection and identification of common bacterial pathogens in cerebrospinal fluid. J Clin Microbiol 38:2076–2080PubMedGoogle Scholar
  18. Maligoy M, Mercade M, Cocaign-Bousquet M, Loubiere P (2008) Transcriptome analysis of Lactococcus lactis in coculture with Saccharomyces cerevisiae. Appl Environ Microbiol 74:485–494PubMedCrossRefGoogle Scholar
  19. Mansy MSM, Fadl AA, Ashour MSE, Khan MI (1999) Amplification of Proteus mirabilis chromosomal DNA using the polymerase chain reaction. Mol Cell Probe 13:133–140CrossRefGoogle Scholar
  20. Masco L, Vanhoutte T, Temmerman R, Swings J, Huys G (2007) Evaluation of real-time PCR targeting the 16 S rRNA and recA genes for the enumeration of bifidobacteria in probiotic products. Int J Food Microbiol 113:351–357PubMedCrossRefGoogle Scholar
  21. Mashsouf RY, Zaman A, Farabani HS (2008) Diagostic multiplex polymerase chain reaction assay for the identification of Pseudomonas aeruginosa from the skin biopsy species in burn wound infections and detection of antibiotic susceptibility. Saudi Med J 29:1109–1114Google Scholar
  22. Mobley H (1996) Urinary tract infections: molecular pathogenesis and clinical management. ASM Press, Washington, DC, pp 245–270Google Scholar
  23. Mobley H, Belas R (1995) Swarming and pathogenicity of Proteus mirabilis in the urinary tract. Trends Microbiol 3:280–284PubMedCrossRefGoogle Scholar
  24. Mobley HL, Island MD, Hausinger RP (1995) Molecular biology of microbial ureases. Microbiol Rev 59:451–480PubMedGoogle Scholar
  25. Mulvaney RL, Bremner JM (1981) Control of urea transformations in soils. Soil Biochem 5:153–196Google Scholar
  26. Nakano S, Kobayashi T, Funabiki K, Matsumura A, Nagao Y, Yamada T (2003) Development of a PCR assay for detection of Enterobacteriaceae in foods. J Food Prot 66:1798–1804PubMedGoogle Scholar
  27. Nakayama J, Hoshiko H, Fukuda M, Tanaka H, Sakamoto N, Tanaka S, Ohue K, Sakai K, Sonomoto K (2007) Molecular monitoring of bacterial community structure in long-aged nukadoko: pickling bed of fermented rice bran dominated by slow-growing lactobacilli. J Biosci Bioeng 104:481–489PubMedCrossRefGoogle Scholar
  28. Penner JL, Hennessy JN (1980) Separate O-grouping schemes for serotyping clinical isolates of Proteus mirabilis and Proteus vulgaris. J Clin Microbiol 12:304–309PubMedGoogle Scholar
  29. Postollec F, Falentin H, Pavan S, Combrisson J, Sohier D (2011) Recent advances in quantitative PCR (qPCR) applications in food microbiology. Food Microbiol 28:848–861PubMedCrossRefGoogle Scholar
  30. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  31. Sauer P, Gallo J, Kesselová M, Kolář M, Koukalová D (2005) Universal primers for detection of common bacterial pathogens causing prosthetic joint infection. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 149:285–288PubMedCrossRefGoogle Scholar
  32. Smith CJ, Osborn AM (2008) Advantages and limitations of quantitative PCR(Q-PCR)-based approaches in microbial ecology. FEMS Microbiol Ecol 67:6–20CrossRefGoogle Scholar
  33. Stankowska D, Kwinkowski M, Kaca W (2008) Quantification of Proteus mirabilis virulence factors and modulation by acylated homoserine lactones. J Microbiol Immunol Infect 41:243–253PubMedGoogle Scholar
  34. Sun K, Hu YH, Zhang XH, Bai FF, Sun L (2009) Identification of vhhP2, a novel genetic marker of Vibrio harveyi, and its application in the quick detection of V. harveyi from animal specimens and environmental samples. J Appl Microbiol 107:1251–1257PubMedCrossRefGoogle Scholar
  35. Suter LS, Ulrich EW, Koelz BS, Street VW (1968) Metabolic variations of Proteus in the Memphis area and other geographical areas. J Appl Microbiol 16:881–889Google Scholar
  36. Takeuchi H, Yamamoto S, Terai A, Kurazono H, Takeda Y, Okada Y, Yoshida O (1996) Detection of Proteus mirabilis urease gene in urinary calculi by polymerase chain reaction. Int J Urol 3:202–206PubMedCrossRefGoogle Scholar
  37. Vanniasinkam T, Lanser JA, Barton MD (1999) PCR for the detection of Campylobacter spp. in clinical specimens. Lett Appl Microbiol 28:52–56PubMedCrossRefGoogle Scholar
  38. Wehrle E, Didier A, Moravek M, Dietrich R, Märtlbauer E (2010) Detection of Bacillus cereus with enteropathogenic potential by multiplex real-time PCR based on SYBR green I. Mol Cell Probes 24:124–130PubMedCrossRefGoogle Scholar
  39. Zhang WW, Sun L (2007) Cloning, characterization, and molecular application of a beta-agarase gene from Vibrio sp. strain V134. Appl Environ Microb 73:2825–2831CrossRefGoogle Scholar
  40. Zhang WW, Sun K, Cheng S, Sun L (2008) Characterization of DegQVh, a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain. Appl Environ Microb 74:6254–6262CrossRefGoogle Scholar
  41. Zhang WW, Chen LX, Liu DY (2011a) Characterization of a marine-isolated mercury-resistant Pseudomonas putida strain SP1 and its potential application in marine mercury reduction. Appl Microbiol Biotechnol 93:1305–1314PubMedCrossRefGoogle Scholar
  42. Zhang WW, Han QX, Liu DY, Chen LX (2011b) Cloning, characterization and molecular analysis of a metalloprotease from Proteus mirabilis. Ann Microbiol 61:757–764CrossRefGoogle Scholar

Copyright information

© Springer-Verlag and the University of Milan 2012

Authors and Affiliations

  • Weiwei Zhang
    • 1
    • 2
  • Zongliang Niu
    • 1
    • 2
  • Kun Yin
    • 1
    • 2
    • 3
  • Ping Liu
    • 1
    • 2
  • Lingxin Chen
    • 1
    • 2
    • 4
    Email author
  1. 1.Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone ResearchChinese Academy of SciencesYantaiPeople’s Republic of China
  2. 2.Shandong Provincial Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone ResearchChinese Academy of SciencesYantaiPeople’s Republic of China
  3. 3.Graduate University of the Chinese Academy of SciencesBeijingChina
  4. 4.Yantai Institute of Coastal Zone ResearchChinese Academy of SciencesYantaiChina

Personalised recommendations