Molecular evaluation of Toxocara species in stray cats using loop-mediated isothermal amplification (lamp) technique as a rapid, sensitive and simple screening assay

Autor: Hamidreza Shokrani, Hamid Azimian, Shirzad Fallahi
Rok vydání: 2020
Předmět:
Zdroj: Veterinary Medicine and Science
Veterinary Medicine and Science, Vol 7, Iss 3, Pp 647-653 (2021)
ISSN: 2053-1095
Popis: Toxocara species are parasitic nematodes of dogs and cats with a worldwide distribution. The adult worm lives in the intestine, and horizontal transmission of the infection occurs through eating paratenic host or embryonated eggs. This study aimed to estimate the molecular prevalence of Toxocara species in stray cats using the loop‐mediated isothermal amplification (LAMP) technique. A total of 95 stool samples were randomly collected from stray cats in Khorramabad city in western Iran. Microscopic examination was performed after the separation and extraction of supernatants. The LAMP reaction was performed using the internal transcribed spacer 2 (ITS2) gene primers of Toxocara species and the appropriate master mix. The overall prevalence of Toxocara spp. in stray cats was 20% (19/95, CI 95%: 0.2 ± 0.08) by parasitological and molecular assessments. The microscopic examination of stool samples revealed that 19 samples were positive for Toxocara. The same 19 positive samples were also positive by the LAMP technique. Interestingly, based on the results of the LAMP assay, out of 95 studied samples, 18 (18.94%; CI 95%: 0.19 ± 0.08) specimens were Toxocara canis, while only 1 (1.05%; CI 95%: 0.005 ± 0.01) sample was diagnosed as Toxocara cati. The relatively high prevalence of Toxocara species in the studied cats shows the role of this species in spreading the parasite and the role of the cats in transmitting this zoonotic parasite. Preventive measures including the control of stray cat's population by castration and protection of public gardens where children play are recommended. The easy, highly sensitive and specific LAMP method is proposed for the differential detection of Toxocara species in animals and humans.
Toxocara species are parasitic nematodes of dogs and cats with a worldwide distribution. Toxocara is a globally soil‐transmitted nematode and human infection has been reported in almost all regions of the world, especially those with tropical weather and abundance of dogs and cats. This parasite is spread not only in developing countries but also in all developed countries so that in some of these countries it is the most common human helminthic infection. Human infections are more commonly reported in children, especially children of low age. These nematodes in humans cause Visceral larva migrans (VLM) syndrome, which is accompanied by different clinical symptoms, sometimes lead to respiratory problems and even death. Toxocara infection can be detected in dogs and cats using history and based on clinical signs. Finding the brown eggs with thick and hollow walls in the fecal specimen determine the diagnosis. The wet mount (direct smear) preparation from the stool specimen solely is not an appropriate method for finding eggs while accompanied by the concentration methods such as flotation assay to make a more accurate diagnosis. The use of new diagnostic methods, such as molecular assays, to determine the rate of infection in dogs and cats as reservoirs of Toxocara, is applicable and can be compared to the results of other tests, including routine stool examination. The loop‐mediated isothermal amplification (LAMP) technique is a simple high‐performance method that was introduced by Notomi et al. (2000), in which the DNA replicated specifically, efficiently, and rapidly under isothermal conditions. The LAMP technique is a simple method that does not require expensive equipment such as Thermal cycler and Gel documentation systems and the reaction can be done in a hot water bath or thermal block. The present study aimed to investigate the molecular frequency of Toxocara species in stray cats in Khorramabad city, Western Iran, using the LAMP technique.
Databáze: OpenAIRE
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