Construction of a urease-negative mutant of Proteus mirabilis: analysis of virulence in a mouse model of ascending urinary tract infection

Proteus mirabilis, a urease-producing uropathogen, causes serious urinary tract infections in humans. To specifically evaluate the contribution of urease to virulence, a mutation was introduced into P. mirabilis HI4320 by homologous recombination. Virulence was assessed in the CBA mouse model of ascending urinary tract infection. Twenty mice each were challenged transurethrally with P. mirabilis HI4320 and its urease-negative derivative (1 x 10(9) to 2 x 10(9) CFU). At 48 h animals were sacrificed and the mean log10 CFU per milliliter of urine (parent, 6.23; mutant, 4.19; P = 0.0014) or per gram of bladder (parent, 6.29; mutant, 4.28; P = 0.0002), left kidney (parent, 4.11; mutant, 1.02; P = 0.00009), and right kidney (parent, 4.11; mutant, 2.43; P = 0.036) were all shown to be significantly different. These data demonstrate a role for urease as a critical virulence determinant for uropathogenic P. mirabilis.

[1]  H. Mobley,et al.  Proteus mirabilis urease: nucleotide sequence determination and comparison with jack bean urease , 1989, Journal of bacteriology.

[2]  H. Mobley,et al.  Proteus mirabilis urease: genetic organization, regulation, and expression of structural genes , 1988, Journal of bacteriology.

[3]  J. Mekalanos,et al.  A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR , 1988, Journal of bacteriology.

[4]  H. Mobley,et al.  Urease-positive bacteriuria and obstruction of long-term urinary catheters , 1987, Journal of clinical microbiology.

[5]  H. Mobley,et al.  Uropathogenicity in rats and mice of Providencia stuartii from long-term catheterized patients. , 1987, The Journal of urology.

[6]  J W Warren,et al.  Fever, bacteremia, and death as complications of bacteriuria in women with long-term urethral catheters. , 1987, The Journal of infectious diseases.

[7]  T. Ahlering,et al.  Hyperammonemic coma due to Proteus infection. , 1985, The Journal of urology.

[8]  J. S. Rodman,et al.  A randomized double-blind study of acetohydroxamic acid in struvite nephrolithiasis. , 1984, The New England journal of medicine.

[9]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[10]  E. T. Johnson The Condom Catheter: Urinary Tract Infection and Other Complications , 1983, Southern medical journal.

[11]  R. Freter,et al.  Ascending, unobstructed urinary tract infection in mice caused by pyelonephritogenic Escherichia coli of human origin , 1983, Infection and immunity.

[12]  A. Pühler,et al.  A Broad Host Range Mobilization System for In Vivo Genetic Engineering: Transposon Mutagenesis in Gram Negative Bacteria , 1983, Bio/Technology.

[13]  R. Adegbola,et al.  Haemagglutinins and fimbriae of Morganella, Proteus and Providencia. , 1982, Journal of medical microbiology.

[14]  D. Musher,et al.  Urease. The primary cause of infection-induced urinary stones. , 1976, Investigative urology.

[15]  D. Musher,et al.  Role of urease in pyelonephritis resulting from urinary tract infection with Proteus. , 1975, The Journal of infectious diseases.

[16]  B. Griffel,et al.  Prevention of ascending pyelonephritis in mice by urease inhibitors. , 1974, Nephron.

[17]  D. MacLaren The influence of acetohydroxamic acid on experimental proteus pyelonephritis. , 1974, Investigative urology.

[18]  N. Carson,et al.  Site of infection in acute urinary-tract infection in general practice. , 1971, Lancet.

[19]  D. Maclaren,et al.  The significance of urease in proteus pyelonephritis: A histological and biochemical study , 1969, The Journal of pathology.

[20]  D. MacLaren,et al.  The significance of urease in proteus pyelonephritis: a bacteriological study. , 1968, The Journal of pathology and bacteriology.

[21]  F. Fitzpatrick Pyelonephritis in the Mouse. I. Infection Experiments , 1966, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[22]  R. H. Gorrill,et al.  THE FATE OF PSEUDOMONAS AERUGINOSA, PROTEUS MIRABILIS AND ESCHERICHIA COLI IN THE MOUSE KIDNEY. , 1965, The Journal of pathology and bacteriology.

[23]  J. Marmur A procedure for the isolation of deoxyribonucleic acid from micro-organisms , 1961 .

[24]  A. Braude,et al.  ROLE OF BACTERIAL UREASE IN EXPERIMENTAL PYELONEPHRITIS , 1960, Journal of bacteriology.

[25]  R. H. Gorrill The effect of obstruction of the ureter on the renal localisation of bacteria. , 1956, The Journal of pathology and bacteriology.

[26]  A. P. Shapiro,et al.  Hematogenous pyelonephritis in rats. I. Its pathogenesis when produced by a simple new method. , 1955, The Journal of clinical investigation.