Isolation and characterization of Listeria monocytogenes from environmental and clinical sources by culture and PCR-RFLP methods

Abstract

Background and Objectives: Due to the widespread distribution of Listeria monocytogenes in environmental and animal sources and serious clinical complications in human, this study was aimed to isolate L. monocytogenes from water and clin- ical specimens by culture and PCR methods and to investigate the presence of hlyA and inlA virulence genes.
 Materials and Methods: Water and clinical samples of vaginal and fecal were screened for the presence of L. monocyto- genes by phenotypic and standard biochemical tests. PCR amplification was performed on extracted DNA using primers based on the hlyA and inlA genes. A 733-bp fragment of inlA gene was used for investigation of polymorphism using RFLP analysis.
 Results: In total, 45 phenotypically and molecularly confirmed L. monocytogenes strains were isolated from different sourc- es including 30 (16.7%) from water, 9 (11.3%) from vaginal swabs and 6 (7.5%) from fecal samples. RFLP analysis of PCR products using AluI and Tsp509I restriction enzymes, generated two profiles with 8 to 10 bands ranging in size from 15 to
 210 bp. The majority of water and clinical isolates were classified in profile 2.
 Conclusion: We demonstrated 45 L. monocytogenes isolates from tested water and clinical samples by phenotypic and mo- lecular tests. The majority of the isolates were classified in the same RFLP profile, showing the water as a potential source of clinical complications in patients in the region of study.

Prevalence and Antimicrobial Resistance Profiles of Foodborne Pathogens Isolated from Dairy Cattle and Poultry Manure Amended Farms in Northeastern Ohio, the United States

“…Confirmation of bacterial spp. for E. coli O157, Salmonella L. monocytogenes , and Campylobacter was performed using a multiplex-PCR assay (mPCR) [ 39 , 42 , 43 , 44 ]. PCR products were visualized using gel-electrophoresis on a 2% agarose gel containing 0.5 µg/mL ethidium bromide.…”

Antimicrobial susceptibility patterns of Listeria monocytogenes isolated from fresh produce in KwaZulu-Natal Province, South Africa

“…In a recent study conducted by Mpondo (2021), it was found that L. monocytogenes was present in 13% (9/69) of water samples collected from river and irrigation waters in the Eastern Cape Province of South Africa. Previously, Meghdadi et al (2019) also detected L. monocytogenes 16.76% (16/180) in river waters. The high prevalence of L. monocytogenes in the river is likely due to anthropogenic inputs (Khatri and Tyagi, 2014).…”

“…Furthermore, the inlA gene, one of L. monocytogenes virulence genes, can be used as a marker for the detection of L. monocytogenes in water and agricultural produce. The inlA gene was recently used for the identification of L. monocytogenes isolated from clinical sources, including vaginal swabs and faeces (Meghdadi et al, 2019).…”

Detection Of Pathogens In Human Consumption Water Through Pcr Technique

“…Nuclease-Free Water was added according to the nmol concentration. Using a 1.5 mL Eppendorf tube, 5 μL of the R primer were added, and it was then filled up with 45 μL of Nuclease-Free Water, resulting in a total solution of 50 μL that recognizes the genus Listeria monocytogenes in 388 bp (15) .…”