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Background  Toxoplasmosis is a widespread zoonotic parasitic disease that occurs in both animals and humans. Traditional molecular assays are often difficult to perform, especially for the early diagnosis of  Toxoplasma gondii  infections. Here, we established a novel loop-mediated isothermal amplification targeting the 529 bp repeat element ( 529 bp -LAMP) to detect  T. gondii  DNA in blood samples of experimental mice infected with tachyzoites of the RH strain.    Findings  The assay was performed with Bst DNA polymerase at 65°C for 1 h. The detection limit of the  529 bp- LAMP assay was as low as 0.6 fg of  T. gondii  DNA. The sensitivity of this assay was 100 and 1000 fold higher than that of the LAMP targeting  B1  gene ( B1 -LAMP) and nested PCR targeting 529 bp repeat element ( 529 bp -nested PCR), respectively. The specificity of the  529 bp- LAMP assay was determined using the DNA samples of  Trypanosoma evansi, Plasmodium falciparum, Paragonimus westermani, Schistosoma japonicum, Fasciola hepatica  and  Angiostrongylus cantonensis . No cross-reactivity with the DNA of any parasites was found. The assay was able to detect  T. gondii  DNA in all mouse blood samples at one day post infection (dpi).    Conclusions  We report the following findings: ( i ) The detection limit of the  529 bp- LAMP assay is 0.6 fg of  T. gondii  DNA; ( ii ) The assay does not involve any cross-reactivity with the DNA of other parasites; ( iii ) This is the first report on the application of the LAMP assay for early diagnosis of toxoplasmosis in blood samples from experimentally infected mice. Due to its simplicity, sensitivity and cost-effectiveness for common use, we suggest that this assay should be used as an early diagnostic tool for health control of toxoplasmosis.

Loop-mediated isothermal amplification (LAMP): Early detection of Toxoplasma gondii infection in mice