Loop mediated isothermal amplification (LAMP) of Toxoplasma DNA from dried blood spots

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

“…However, both PCR and LAMP failed to detect T. gondii in blood samples 56 days post-infection. In addition, both LAMP and routine PCR, targeting the 529 bp RE gene, did not detect T. gondii in DNA extracted from blood of mice during the chronic phase of the disease, which can be attributed to progressive decrease in parasitemia levels as infection continues [10]. Taken together, these findings indicate that LAMP-based detection of toxoplasma is limited to the early stages of infection and is difficult to detect across peripheral blood at the quiescent bradyzoite stage.…”

“…The molecular detection of T. gondii is not only limited by the detection method, but is also related to the stage of infection of the animals to be tested. Hegazy et al compared detection rates of T. gondii in blood samples of mice at different stages after infection with an ME49 strain [10]. According to results, LAMP revealed 18 positives, out of the 20 examined samples, on the seventh day post-infection.…”

“…Particularly, the serum IgM/IgG antibody test has been widely used as a primary screening method for toxoplasmosis infection. However, this method may fail to detect specific anti-T. gondii antibodies during the active infection phase in animals, because these antibodies may only be produced after several weeks of parasitemia [10]. In recent years, PCR, realtime PCR, and nested-PCR assays have become essential tools for the molecular diagnosis of T. gondii.…”

“…LAMP products can be confirmed by the turbidity of the resulting magnesium pyrophosphate by visual inspection, or addition of a fluorescent dye to the reaction system which makes the product visible under UV light [27]. LAMP has several advantages over traditional PCR, hence it is at the forefront of research in the search for new diagnostic tools for parasitic diseases [10]. During toxoplasmosis diagnosis, a series of LAMP-based assays, targeting B1 gene or 529 bp repeat sequences, internal transcriptional spacer sequences (ITS-1), as well as 18S rDNA sequences, have been established.…”

A Novel Rapid Visual Detection of Toxoplasma Gondii by Combining Recombinase Polymerase Amplification and Lateral Flow Dipstick Coupled With CRISPR-Cas13a Fluorescence Assay

“…However, pathologic tissue examination or strain isolation are mostly used for diagnosis of animal infection, less frequently for that of human toxoplasmosis [1]. Currently, nucleic acid and speci c antibody assays of T. gondii are the most commonly used techniques in clinical settings [9]. Particularly, the serum IgM/IgG antibody test has been widely used as a primary screening method for toxoplasmosis infection.…”