A fast and easy detection for Listeria monocytogenes in urine of women using polymerase chain reaction and fluorescent probes hlyA gene

Subject Areas : Diagnostic Microbiology

Mohammad Sbhani Lari ¹ , Nader Shahrokhi ² , Mohammad Kargar ³

1 - MS.c., Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.
2 - Associate Professor, Department of Molecular Biology, Microbiology Group, Pasteur Institute, Tehran, Iran.
3 - Associate Professor, Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.

Received: 2014-03-14 Accepted : 2014-06-01 Published : 2014-10-01

Keywords: Urine, Listeria, Fluorescent probe,

Abstract :

Background & Objectives: Listeria monocytogenes is one of the etiologic causes of asymptomatic infection that can cause urogenital tract infection in infants and fetal death. Given the challenges posed by growing the bacteria, this study aimed to detect Listeria in urine using polymerase chain reaction and fluorescent probes. Materials & Methods: This cross-sectional study was carried out on 100 urine samples obtained from the patients who were referred to a specialized center for women in Jahrom and Larestan. The sensitivity of the technique was evaluated using a standard strain. Then hlyA gene probes were designed using the polymerase chain reaction, and fluorescence level was determined by fluorescent probes. Results: Based on FLASH-PCR assay between 5 to 10 genes were detected in each reaction. Using fluorescent probes for L. monocytogenes hlyA gene overall, 30 out of 100 clinical samples (30%) were infected to this bacterium. Conclusion: The results showed that fluorescent probe increase speed, sensitivity and specificity of the molecular identification of Listeria and therefore it can be a suitable alternative to electrophoresis. Thus, with respect to the accuracy of this non-invasive method it is possible to monitor larger amounts of samples suspected to L. monocytogenes infections.