Abstract
Leptospirosis is a zoonosis prevalent in tropical countries and affects animals and humans alike. Leptospira interrogans, the causative organism for this waterborne infection, spreads through the urine of infected animals. There is a direct link between contaminated water and Leptospira outbreaks. This study reports a rapid assay to detect and differentiate pathogenic Leptospira from non-pathogenic in environmental water using multiplex PCR. The assay uses primers targeting the Lipl32 and Lipl21 gene. The multiplex PCR has been standardized using 11 pathogenic and one saprophytic serovar of Leptospira. The analytical sensitivity of the developed method was evaluated with different concentrations of template DNA. This method was used to screen water samples collected from 20 different sources from Chengalpattu town in Kancheepuram District, Tamil Nadu, India. Of the 20 water samples screened, 13 samples tested positive for pathogenic Leptospira, and seven samples tested negative. Four water samples were found to carry both pathogenic and saprophytic species. The developed multiplex PCR assay is highly useful for detecting and distinguishing pathogenic and saprophytic leptospires in water.
Highlights
Leptospirosis is a zoonosis having a global presence with a higher prevalence in tropical regions
This study reports a rapid assay to detect and differentiate pathogenic Leptospira from non-pathogenic in environmental water using multiplex PCR
Of the 20 water samples screened, 13 samples tested positive for pathogenic Leptospira, and seven samples tested negative
Summary
Leptospirosis is a zoonosis having a global presence with a higher prevalence in tropical regions. Heavy rainfall and flooding result in large epidemics causing deaths in developed and developing countries[1,2,3]. Humans are infected via exposure to the urine of infected animals or through contaminated soil or water 4. Leptospira species display different affinities and adaptations in different hosts and exhibit clinical symptoms of varying degrees[5]. Rodents are reservoirs and carriers of Leptospira but rarely show symptoms of the disease. Other animals like livestock, cattle, and pets exhibit severe symptoms, including multiple organ injury and abortion upon infection[6,7]. In infected humans, the disease’s clinical manifestations range from mild, flu-like symptoms to severe conditions like jaundice, renal impairment, and hemorrhage to life-threatening conditions[5]. A direct correlation between Leptospira epidemics and direct contact with contaminated environmental water has been reported. A damaged skin barrier like a cut or an open wound is the most common route of pathogen entry[2,8,9]
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