title: Molecular detection of HpmA and HlyA hemolysin of uropathogenic Proteus mirabilis.
abstract: Urinary tract infection (UTI) is one of the bacterial infections frequently documented in humans. Proteus mirabilis is associated with UTI mainly in individuals with urinary tract abnormality or related with vesicular catheterism and it can be difficult to treat because of the formation of stones in the bladder and kidneys. These stones are formed due to the presence of urease synthesized by the bacteria. Another important factor is that P. mirabilis produces hemolysin HpmA, used by the bacteria to damage the kidney tissues. Proteus spp. samples can also express HlyA hemolysin, similar to that found in Escherichia coli. A total of 211 uropathogenic P. mirabilis isolates were analyzed to detect the presence of the hpmA and hpmB genes by the techniques of polymerase chain reaction (PCR) and dot blot and hlyA by PCR. The hpmA and hpmB genes were expressed by the RT-PCR technique and two P. mirabilis isolates were sequenced for the hpmA and hpmB genes. The presence of the hpmA and hpmB genes was confirmed by PCR in 205 (97.15 %) of the 211 isolates. The dot blot confirmed the presence of the hpmA and hpmB genes in the isolates that did not amplify in the PCR. None of the isolates studied presented the hlyA gene. The hpmA and hpmB genes that were sequenced presented 98 % identity with the same genes of the HI4320 P. mirabilis sample. This study showed that the PCR technique has good sensitivity for detecting the hpmA and hpmB genes of P. mirabilis.
authors.affiliation: Laboratório de Bacteriologia, Departamento de Microbiologia, Universidade Estadual de Londrina (UEL), Cx. P. 6001, Londrina, PR, 86051-970, Brazil.
authors.forename: Silvia Emanoele
authors.initials: SE
authors.lastname: Cestari
authors.forename: Marilucia Santos
authors.initials: MS
authors.lastname: Ludovico
authors.forename: Fernando Henrique
authors.initials: FH
authors.lastname: Martins
authors.forename: Sérgio Paulo Dejato
authors.initials: SP
authors.lastname: da Rocha
authors.forename: Waldir Pereira
authors.initials: WP
authors.lastname: Elias
authors.forename: Jacinta Sanchez
authors.initials: JS
authors.lastname: Pelayo
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics*; D015139:Blotting, Southern; D029968:Escherichia coli Proteins / Q000235:genetics; D006460:Hemolysin Proteins / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011513:Proteus mirabilis / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014552:Urinary Tract Infections / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; C064194:HpmA hemolysin; C104106:HpmB protein, Proteus mirabilis; D008565:Membrane Proteins