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Quick Identification and Differentiation of Proteus spp. by PCR and RFLP Along with Their Antibiotic Resistance
H Jannat1, S M Shamsuzzaman, M A Faisal
1Dr Hasnatul Jannat, Assistant Surgeon, Mymensingh Medical College Hospital, Mymensingh, Bangladesh;
Abstract:
Different Proteus species are encountered in human infections and may vary with the type of infections they cause. So, the present study was conducted to detect species of Proteus by PCR and RFLP along with their antibiotic resistance pattern. This cross-sectional study was conducted in the Department of Microbiology of Dhaka Medical College, Dhaka, Bangladesh, from July 2018 to June 2019. A total of 500 wound swab and pus, urine and blood samples were tested for bacterial pathogens. Proteus spp. were identified and differentiated by biochemical test, PCR and RFLP. Antibiotic susceptibility was performed by disc-diffusion technique. Fourty Proteus spp. was isolated from 300 culture positive samples, giving 13.33% prevalence of Proteus infections. Proteus mirabilis and Proteus vulgaris were identified by culture, biochemical test, PCR and RFLP. The results were similar by both methods (biochemical tests and PCR). RFLP of 16S rRNA fragments digested with HaeIII revealed that P. mirabilis consisted of two bands at approximately 110 and 190 bp and P. vulgaris consisted of three bands at approximately 100, 180 and 220 bp. The proportion (80%) of P. mirabilis was more than P. vulgaris. Highest proportion (77.5%) of Proteus spp. was isolated from wound swab and pus followed by urine samples. A significant proportion of Proteus spp. was multidrug resistant (90%) and extensively drug resistant (37.5%). Fosfomycin was found the most sensitive drug followed by imipenem. This study provided an insight into antibiotic resistance pattern of Proteus spp. and showed high level resistance towards commonly used antimicrobial agents. PCR and RFLP may be suitable method to identify and differentiate species of Proteus and to treat them accordingly.
Insights
This study identified Proteus mirabilis and Proteus vulgaris in infections using PCR and RFLP, revealing high multidrug resistance. Fosfomycin and imipenem showed the most sensitivity against these Proteus species.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Proteus species are significant causes of human infections.
- Accurate species identification is crucial for effective treatment.
- Understanding antibiotic resistance patterns is vital for public health.
Purpose of the Study:
- To identify Proteus species using Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP).
- To determine the antibiotic resistance patterns of isolated Proteus species.
- To evaluate the efficacy of PCR and RFLP in differentiating Proteus species.
Main Methods:
- Isolation and identification of Proteus species from wound, urine, and blood samples.
- Species differentiation using biochemical tests, PCR, and RFLP of 16S rRNA gene fragments.
- Antibiotic susceptibility testing via disc-diffusion method.
Main Results:
- Proteus mirabilis and Proteus vulgaris were identified, with P. mirabilis being more prevalent (80%).
- Highest isolation rate was from wound swabs and pus (77.5%), followed by urine samples.
- A high prevalence of multidrug resistance (90%) and extensively drug resistance (37.5%) was observed.
- Fosfomycin and imipenem demonstrated the highest sensitivity.
Conclusions:
- PCR and RFLP are effective methods for identifying and differentiating Proteus species.
- Proteus infections exhibit significant multidrug resistance, necessitating careful antibiotic selection.
- Fosfomycin and imipenem are potential treatment options for Proteus infections.
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