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Tandem tetramer-based microsatellite fingerprinting for typing of Proteus mirabilis strains
Tomasz Cieślikowski1, Dobrosława Gradecka, Magdalena Mielczarek
1Centre of Microbiology and Virology, Polish Academy of Sciences, Łódź, Poland. tcieslik@cmiwpan.lodz.pl
Abstract:
Two microsatellite tandem repeated tetramers, (GACA)(4) and (CAAT)(4), were used for Proteus mirabilis strain differentiation. The microsatellite-based PCR tests were applied for the examination of interstrain diversity for 87 P. mirabilis strains. Forty-six of the investigated strains were clinical isolates (5 were hospital isolates and 39 were outpatient clinic isolates); 42 strains were derived from the Kauffmann-Perch collection of laboratory strains. Fingerprinting done with the tetramers had a high discrimination ability [0.992 and 0.940 for (GACA)(4) and (CAAT)(4), respectively]. The distributions of clinical isolates among well-defined laboratory strains, determined by numerical analysis (unweighted pair-group method with arithmetic averages; Dice similarity coefficient), proved their genetic similarity to reference strains in the Kauffmann-Perch collection. This analysis also indicated that it is possible to estimate some phenotypic properties of P. mirabilis clinical isolates solely on the basis of microsatellite fingerprinting.
Insights
This study differentiated Proteus mirabilis strains using microsatellite PCR. The methods revealed high discrimination ability, linking clinical isolates to laboratory strains and suggesting phenotypic property prediction from genetic fingerprints.
Area of Science:
- Microbiology
- Genetics
Background:
- Proteus mirabilis is an opportunistic pathogen.
- Strain differentiation is crucial for understanding its epidemiology and pathogenesis.
- Microsatellite markers offer a high-resolution typing method.
Purpose of the Study:
- To evaluate the utility of two tetrameric microsatellite markers, (GACA)(4) and (CAAT)(4), for differentiating Proteus mirabilis strains.
- To assess the interstrain diversity within a collection of clinical and laboratory P. mirabilis isolates.
- To explore the potential of microsatellite fingerprinting for predicting phenotypic characteristics.
Main Methods:
- Polymerase chain reaction (PCR) amplification using two specific microsatellite primers: (GACA)(4) and (CAAT)(4).
- Analysis of 87 P. mirabilis strains, including 46 clinical isolates and 42 laboratory strains from the Kauffmann-Perch collection.
- Numerical analysis using the unweighted pair-group method with arithmetic averages (UPGMA) and the Dice similarity coefficient for genetic similarity assessment.
Main Results:
- Both (GACA)(4) and (CAAT)(4) microsatellites demonstrated high discriminatory power (0.992 and 0.940, respectively).
- Genetic analysis revealed that clinical isolates were genetically similar to reference strains within the Kauffmann-Perch collection.
- The study successfully differentiated between various P. mirabilis strains based on their microsatellite profiles.
Conclusions:
- Microsatellite-based PCR using (GACA)(4) and (CAAT)(4) is a powerful tool for P. mirabilis strain differentiation and diversity assessment.
- The findings support the genetic relatedness of clinical P. mirabilis isolates to established laboratory strains.
- Microsatellite fingerprinting holds promise for inferring phenotypic traits of P. mirabilis clinical isolates, aiding in epidemiological studies.