Molecular Diagnosis of Clinically Isolated <i>Klebsiella pneumoniae</i> Strains by PCR-ELISA

Abstract

<em>Klebsiella pneumoniae</em> is the most important infectious bacteria in Enterobacteriaceae family and the most common bacteria causing Urinary Tract Infection (UTI) after <em>Escherichia coli</em>. Therefore, accurate and rapid identification of this bacterium in hospital infection is very important.In this study, PCR-ELISA method was used for detecting <em>Klebsiella pneumonia </em>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 <em>Klebsiella pneumoniae</em> 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<em> Klebsiella pneumoniae </em>strains.