Molecular Diagnosis of Clinically Isolated Klebsiella pneumoniae Strains by PCR-ELISA

Klebsiella pneumoniae is the most important infectious bacteria in Enterobacteriaceae family and the most common bacteria causing Urinary Tract Infection (UTI) after Escherichia coli. Therefore, accurate and rapid identification of this bacterium in hospital infection is very important.In this study, PCR-ELISA method was used for detecting Klebsiella pneumonia clinical strains. For this purpose, 16S rDNA gene based specific primers were designed and the DIG-labeled PCR products were bound to streptoavidin-coated wells of a microtiter plate and detected by anti-DIG–peroxidase conjugate. Biotin-labeled DNA probe specific for 16S rDNA gene was used in PCR-ELISA.Sensitivity and specificity of PCR-ELISA method were determined by using Enterobacteria strains. 16S rDNA of Klebsiella pneumoniae was amplified using gene specific primers resulted in a fragment of the 260 bp. The results of PCR-ELISA showed that this technique does not cross-react with the bacteria in their families as well as the sensitivity of 6.0 ng were evaluated.PCR-ELISA is known as an accurate and rapid method for detection of the infectious agents and therefore can be used as a suitable substitute for all the above aspects because it is quite a sensitive, specific, and rapid method for detection of the Klebsiella pneumoniae strains.

Keywords:

Klebsiella pneumoniae , 16S rDNA , Diagnostic Method , PCR-ELISA