Establishment of a PCR-based diagnostic method for the identification of Klebsiella pneumoniae

3^rd International Conference on Diagnostic Microbiology and Infectious Diseases

September 24-25, 2018 | Montreal, Canada

AA Bastawy, HM El-Refaie, YA Ramadan, R Wasfi and M Tawfick

October University for Modern, Egypt

Posters & Accepted Abstracts: J Med Microb Diagn

Abstract :

Klebsiella pneumoniae is one of the significant causative agents for nosocomial infections. The rate of infections by this bacterium has increased dramatically and thus searching for quick and sensitive molecular methods for its detection became a must. Our research aims at validating a sensitive and specific PCR based method for the diagnosis of infections caused by Klebsiella pneumoniae. Our work started by using online bioinformatics tools to find a unique gene that can be used for detection of Klebsiella pneumoniae and design specific primer for its amplification. In silico studies results in choosing the rcsA gene, regulating colanic acid capsular biosynthesis activation protein A, as a unique gene for Klebsiella pneumoniae. A primer pair (named K1) was designed to target the selected gene, followed by determining primers sensitivity and specificity in Klebsiella pneumoniae detection. The designed primer was able to amplify targeted gene sequence in genomic DNA extracted from standard and clinical strains and in diluted DNA template concentration up to 10-1ng/μl. K1 primer pair worked well bacterial culture without DNA extraction with high sensitivity reaching 10CFU/ml. designed primer failed to amplify genomic DNA template of common Gram-negative uropathogens including Klebsiella oxytoca, Pseudomonas aeruginosa, Escherichia coli and Proteus mirabilis, thus indicating its specificity. In conclusion, the rcsA gene is a unique gene that can be used for sensitive and specific detection of Klebsiella pneumoniae.

Biography :

E-mail: a.a.bastawy@gmail.com