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