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Molecular Detection of HpmA and HlyA Hemolysin of Uropathogenic Proteus mirabilis
Authors:Silvia Emanoele Cestari  Marilucia Santos Ludovico  Fernando Henrique Martins  Sérgio Paulo Dejato da Rocha  Waldir Pereira Elias  Jacinta Sanchez Pelayo
Institution:1. Laboratório de Bacteriologia, Departamento de Microbiologia, Universidade Estadual de Londrina (UEL), Cx. P. 6001, Londrina, PR, 86051-970, Brazil
2. Laboratório de Bacteriologia, Instituto Butantan, S?o Paulo, SP, Brazil
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.
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