Evaluation on the use of β-lactamase and
Aminoglycoside modifying enzyme gene sequences as markers for the early
detection of antibiotic resistance profile of Pseudomonas
aeruginosa
Affiliations: Department of Biochemistry, PSG College of Arts and
Science, Coimbatore, India | Department of Microbiology, PSG Institute of Medical
Sciences and Research, Coimbatore, India
Note: [] Corresponding author: Lecturer, Department of Biochemistry, PSG
College of Arts and Science, Coimbatore, Pin – 641 014, India. Tel.: +91 422
2220232; Fax: +91 422 2575622; E-mail; [email protected]
Abstract: Pseudomonas aeruginosa is one of the major causes of infections
including the hospital acquired (Nosocomial) infections. Detection of them and
their antibiotic resistance profile by conventional method takes about three
days. Recently, DNA based diagnostic methods are being used for the
identification of the pathogens. Hence we have tested a rapid and sensitive
method using DNA sequences as markers for detecting the presence of three genes
coding for the enzymes that inactivate the two most commonly used
Anti-pseudomonadal drugs such as β-lactam antibiotics (Penicillin, and
its derivatives) and Aminoglycosides such as Gentamicin, Tobramycin, Amikacin,
Streptomycin. The internal region of these genes were used for designing and
synthesizing primers and these primers were used in Polymerase Chain Reaction
(PCR) to screen for the presence of these genes in the clinical isolates and to
label them non-radioactively with Biotin. They in turn were used to detect the
presence of the antibiotic resistance genes in the clinical isolates by
hybridization. The specificity (ratio of positive results obtained in both
methods and the sensitivity (the minimum amount of sample DNA and the labeled
probe required for the tests) were evaluated.
Keywords: gene marker, DNA probes, antibiotic resistance, drug-modifying enzymes, non-radioactive labelling, anti-biogram, culture and sensitivity test
