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

Abstract

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× 10 3 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.

Association among biofilm formation, virulence gene expression, and antibiotic resistance in Proteus mirabilis isolates from diarrhetic animals in northeast China

“…In this study, 90.91% of the isolates were positive for ureC. However, its prevalence was relatively low when compared to the results of an earlier study [30]. Our findings revealed that testing only ureC increased the likelihood of obtaining P. mirabilis negative results.…”

“…As a target gene, ureC, was used to positively identify P. mirabilis as described previously [30]. Ali et al.…”

Association among biofilm formation, virulence gene expression, and antibiotic resistance in Proteus mirabilis isolates from diarrhetic animals in northeast China

“…In this study, 90.91% of the isolates were positive for ureC. However, its prevalence was relatively low when compared to the results of an earlier study [30]. Our results revealed that only testing ureC geneto prove that P. mirabilis may be get negative results.…”

“…As a target gene, ureC, was used to positively identify P. mirabilis as described previously [30]. Ali et al reported that 96.66% of human P. mirabilis isolates (n = 30) recovered from the urinary tract expressed ureC [29].…”

Genomes of gut bacteria fromNasoniawasps shed light on phylosymbiosis and microbe-assisted hybrid breakdown

“…To quantitatively determine the co-culture composition of the Proteus mirabilis and Providencia rettgeri biofilm community we developed a qPCR assay using primers specific for unique single-copy genes in each respective genome for P. mirabilis (5’-GGTGAGATTTGTATTAATGG and 5’-ATCAGGAAGATGACGAG, annealing temperature 58°C) and P. rettgeri (5’-AACTCGGTCAGTTCCAAACG and 5’-CTGCATTGTTCGCTTCTCAC, annealing temperature 66°C). Proteus primers were designed for the ureR gene using a previously reported forward primer [123] and a new reverse primer. Providencia primers were designed based on a phage gene only found within the Providencia genomes.…”