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 ◽  
...  

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.

Parasite ◽  
2021 ◽  
Vol 28 ◽  
pp. 41
Author(s):  
Yangji Xue ◽  
Qingming Kong ◽  
Haojie Ding ◽  
Chengzuo Xie ◽  
Bin Zheng ◽  
...  

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.


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