Study of Urease gene and O-Antigen Cluster Gene of Proteus Mirabilis Isolated from Urinary Stones Patients

The aims form this study were to determine the serotypes of Proteus mirabilis isolated from urinary stone patients by using Nuclear magnetic resonance (NMR) spectroscopy and genotype characterizations of UreR gene as well as O-antigen cluster of these isolates via polymerase chain reaction (PCR) technique and sequences alignment process of these genes. The samples were collected from urinary stone patients, bacterial isolates were identified by traditional methods, Vitek 2 system technique and PCR technique, DNA samples were extracted according to the manufacture kit. The lipopolysaccharide and O-antigen of bacterial isolates were isolated, extracted, purified and electrophoresed. The Serotyping of bacterial isolates was determined by using NMR spectroscopy. The genotype study was included two steps, first the detection of UreR gene and O-antigen by PCR technique and seconds the determination the sequencing of these two genes according to Sanger and Coulson method. The determination of Proteus mirabilis serogroups showed that four different types of serogroups for the tested isolates, which included O3, O16, O18, and O20. These serogroups were tested by Nuclear magnetic resonance (NMR) spectroscopy via the determination of the chemical structure of O-antigen repeating units of tested isolates. The detection of the UreR gene and O-antigen by using polymerase chain reaction (PCR) showed that all the tested isolates of P. mirabilis possessed the two genes, where the produced band of UreR gene and O-antigen was 405 bp and 342 bp, which represents the presence of the UreR gene and O-antigen, respectively. The sequencing and the constructed phylogenetic tree results of UreR gene and O-antigen showed that the P. mirabilis isolates were generally closely related to their respective type. NMR spectroscopy was considered a suitable method for determine the serogroup of P. mirabilis based on the determination of chemical structure of O-antigen repeating units. The UreR and O-antigen gene sequence comparison seems to be an appropriate method for inferring genetic relationships within the P. mirabilis isolates on a molecular basis. The UreR gene and O-antigen gene cluster sequences of P. mirabilis isolates allow rapidly a specific PCR assays to be designed.