Rapid Real Time PCR Based Detection of Cell Count in Case of Urinary Tract Infection

Abstract

Microbial identification and antimicrobial susceptibility testing methods currently used in clinical microbiology laboratories require at least two to three days because they rely on the growth and isolation of micro-organisms. This long, but necessary, delay has enormous consequences on prophylactic usage of antimicrobial drugs. This study was an attempt to reduce this detection time span. Taq Man Real Time PCR has been used as an important tool in the differentiation of Gram nature of bacteria present in UTI patients that allows detection of spiked bacterial 16S rDNA from urine samples within a short span of 5h and also gives us the corresponding cell count of both/either Gram positive and negative organisms present. A standard curve was generated which was used to determine the cell count of control as well as patient samples. Detection could be done in the range of 10³ to 10⁶ cells/mL Patient samples screened clustered either in the allele 1 or allele 2 axes, depending on majority concentration of Gram nature of the micro-organisms. The cell counts for control individuals were scattered within 0 to 10², while very few in the range of 10⁴. The case was just reverse for patient group, where most of the points were scattered within 10⁴ to 10⁸. Thus the optimal selection of appropriate antimicrobials (depending on the gram nature) by clinicians, will be gradually improved as an increasing number of rapid molecular diagnostic tools for the detection, identification and characterization of infectious agents become commercially available.