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

2021 ◽  
Author(s):  
Hamid Azimian ◽  
Hamidreza Shokrani ◽  
Shirzad Fallahi
Keyword(s):  
Isothermal Amplification ◽  
Screening Assay ◽  
Loop Mediated Isothermal Amplification ◽  
Stray Cats ◽  
Simple Screening ◽  
Toxocara Species ◽  
Molecular Evaluation

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

2020 ◽  
Author(s):  
Hamid Azimian ◽  
Hamidreza Shokrani ◽  
Shirzad Fallahi
Keyword(s):  
Microscopic Examination ◽  
Clinical Symptoms ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Larva Migrans ◽  
Parasitic Nematodes ◽  
Screening Assay ◽  
Loop Mediated Isothermal Amplification ◽  
Stray Cats ◽  
Stool Samples

Abstract Background: Toxocara species are parasitic nematodes of dogs and cats with a worldwide distribution. The adult worm lives in the intestine of dogs and cats, and infection occurs through eating paratenic host or embryonated eggs. These nematodes are considered as one of the common pathogens of humans and livestock, and in humans cause Visceral larva migrans (VLM) syndrome, which is accompanied by different clinical symptoms, sometimes lead to respiratory problems and even death. The present study aimed to investigate the molecular frequency of Toxocara species in stray cats using the loop-mediated isothermal amplification (LAMP) technique. Methods: During 3 months (July to September 2018) a total of 95 stool samples were randomly collected from stray cats in Khorramabad city, Lorestan province, West of Iran. To separate the parasite’s eggs, the flotation method with chlorine solution was applied to saturation. Microscopic examination was performed after the separation and extraction of supernatants. To rupture the four layers of Toxocara egg, and extraction of DNA, sonication and freeze and thaw processes were used. After DNA extraction from the stool specimens, the LAMP reaction was performed using the internal transcribed spacer 2 (ITS2) gene primers of Toxocara species and the appropriate LAMP master mix. Results: The overall prevalence of Toxocara spp. in stray cats of Khoramabad city was 20% (19/95) by parasitological and molecular assessments. The microscopic examination of stool samples revealed that 19 samples were positive for Toxocara spp. The same 19 positive samples were also positive in the LAMP experiments. Interestingly, based on the results of the LAMP assay, out of 95 studied samples, 18 (18.41%) specimens were belonged to Toxocara canis, while only 1 (0.5%) sample was diagnosed as Toxocara cati. Conclusion: Identification of Toxocara spp. in dogs and cats as the important sources of human infections especially for children is particularly important. The relatively high prevalence of Toxocara species, especially and interestingly the canis species in the studied cats shows the role of these animals in spreading the disease and the risk of transmission of infection to humans. Precautionary measures such as preventing cats from traveling in public places, especially children's playgrounds, as well as avoiding direct contact with stray cats are recommended.

Rapid Detection of Toxoplasma gondii: Prevalence Investigation of T. gondii Infection Among Stray Cats and Dogs in Zhejiang Province Based on a New Loop-Mediated Isothermal Amplification-Lateral-Flow-Dipstick (LAMP-LFD) Device

2020 ◽  
Author(s):  
Yangji Xue ◽  
Shaohong Lu ◽  
Qingming Kong ◽  
Haojie Ding ◽  
Jianzu Ding ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Lateral Flow ◽  
Zhejiang Province ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Infection Prevalence ◽  
Loop Mediated Isothermal Amplification ◽  
Stray Cats ◽  
Lateral Flow Dipstick ◽  
Parasitic Pathogens

Abstract Background: Toxoplasma gondii (T. gondii) is worldwide spread caused Toxoplasmosis threatening warm-blooded animal and human health, especially for immunodeficient population and pregnant women. Simple and applicable diagnostic methods are urgently needed for the prevention of toxoplasmosis. The molecular diagnosis of T. gondii infection generally requires high technical skills, sophisticated equipment and a controlled lab environment.Methods: In this study, we developed a loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) assay that specifically targets the 529 bp for the detection of T. gondii infection in a new kind of portable device, which is universal, fast, user-friendliness, experimental sensitivity and low risk of aerosol contamination. Results: The detection limit of the LAMP-LFD assay is 1 fg of T. gondii DNA and no cross-reaction with other parasitic pathogens including Leishmania donovani, Plasmodium vivax, Cryptosporidium parvum, etc. In total, 318 stray cat and dog blood samples were collected from Deqing, Wenzhou, Yiwu, Lishui and Zhoushan cities in Zhejiang province, Eastern China. The current infection prevalence of T. gondii was 4.76% and 4.69% in stray cats and dogs respectively, detected by LAMP-LFD device.Conclusions: In conclusion, the established LAMP-LFD was an efficient and avoidable aerosol-contaminated device that can detect 1 fg genomic DNA of T. gondii, and suitable for T. gondii detection in the basic medical institution and even in field areas.

scholarly journals A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs

Parasite ◽  
2021 ◽  
Vol 28 ◽  
pp. 41
Author(s):  
Yangji Xue ◽  
Qingming Kong ◽  
Haojie Ding ◽  
Chengzuo Xie ◽  
Bin Zheng ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Diagnostic Tools ◽  
Loop Mediated Isothermal Amplification ◽  
Stray Cats ◽  
Lateral Flow Dipstick ◽  
Parasitic Pathogens

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis and threatens warm-blooded animal and human health worldwide. Simple and applicable diagnostic methods are urgently needed to guide development of effective approaches for prevention of toxoplasmosis. Most molecular diagnostic tools for T. gondii infection require high technical skills, sophisticated equipment, and a controlled lab environment. In this study, we developed a loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) assay that specifically targets the 529 bp for detecting T. gondii infection. This novel portable device is universal, fast, user-friendly, and guarantees experimental sensitivity as well as low risk of aerosol contamination. Our LAMP-LFD assay has a detection limit of 1 fg of T. gondii DNA, and shows no cross-reaction with other parasitic pathogens, including Cryptosporidium parvum, Leishmania donovani, and Plasmodium vivax. We validated the developed assay by detecting T. gondii in DNA extracted from blood samples collected from 318 stray cats and dogs sampled from Deqing, Wenzhou, Yiwu, Lishui and Zhoushan cities across Zhejiang province, Eastern China. The LAMP-LFD device detected T. gondii DNA in 4.76 and 4.69% of stray cats and dogs, respectively. In conclusion, the developed LAMP-LFD assay is efficient, minimizes aerosol contamination, and is therefore suitable for detecting T. gondii across basic medical institutions and field settings.

scholarly journals Comparison of Conventional PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Arcobacter Species

2013 ◽  
Vol 52 (2) ◽  
pp. 557-563 ◽  
Author(s):  
X. Wang ◽  
D. J. Seo ◽  
M. H. Lee ◽  
C. Choi ◽  
A. B. Onderdonk
Keyword(s):  
Multiplex Pcr ◽  
Rapid Detection ◽  
Isothermal Amplification ◽  
Conventional Pcr ◽  
Loop Mediated Isothermal Amplification ◽  
Pcr Multiplex
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