Additional file 1 of Application of gold immunochromatographic assay strip combined with digital evaluation for early detection of Toxoplasma gondii infection in multiple species

Additional file1: Table S1. PCR primers for the construction of recombinant plasmids. Table S2. Different concentrations of AMA1 protein variants tested using ELISA with serum from four rabbits. Table S3. Test results of 15 serum-positive cat samples using AMA1C-GICA strips with HMREADER. Table S4. Test results of 14 serum-positive different animal samples using AMA1C-GICA strips with HMREADER. Table S5. Test results of four serum-positive human samples using AMA1C-GICA strips with HMREADER

Primers used for the generation of the recombinant proteins.

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Summary of Proteins identified of serum-derived exosomes.

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The dynamic changes of mouse proteins in serum-derived exosomes in response to <i>E</i>. <i>multilocularis</i> infection.

(A) and (B) Transmission electron microscopy of the enriched extracellular vesicles (EVs) isolated from the sera of E. multilocularis-infected (A) and uninfected mice (B). (C) The diameter distribution of the purified EVs. (D) Western blotting analysis of mammalian EV-marker CD63 in the purified EVs. (E) Heatmap of differentially abundance proteins between the serum exosomes from E. multilocularis-infected and uninfected mice. (F) Western blotting analysis the abundance of TGF-β1, GPATC8, LRP1, and AMY1 during E. multilocularis infection.</p

Parasite-origin protein components in serum-derived exosomes in response to <i>E</i>. <i>multilocularis</i> infection.

(A) Immune electron microscopy of EVs isolated from the sera of E. multilocularis-infected and uninfected mice using anti-Em14-3-3 antibodies. The red arrows indicated the parasite-derived EVs. (B) Western blotting analysis of TPx-1, TER ATPase, and FBPA in the serum-derived exosomes at 30-, 60-, 90-day post infection. The uninfected mouse sera were used as control. (C) Immunofluorescence localization of TER ATPase and TPx-1 in the E. multilocularis-infected mouse liver. CW, cyst wall; GL, germinal layer; HF, hydatid fluid; LL, laminated layer; ps, protoscolexs; liver, mouse liver. The nucleus was stained by DAPI. Localization of TER ATPase or TPx-1 was indicated by yellow arrows. (D) The recombinant TPx-1 and TER ATPase could recognize by E. multilocularis-positive sera at 30-, 60, 90-day post infection. The pre-infection sera (at 0-day post infection) were used as control.</p

The performance of TPx-1 and TER ATPase in diagnosis of animal echinococcosis.

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Summary of parasite proteins in serum-derived exosomes from <i>E</i>. <i>multilocularis</i>-infected mice.

Summary of parasite proteins in serum-derived exosomes from E. multilocularis-infected mice.</p

Preparation of polyclonal antibodies against recombinant TPx-1 or TER ATPase.

(A) and (B) SDS-PAGE analysis of purified recombinant TPx-1 or TER ATPase (truncated).(C) Western blotting analysis of native TPx-1 using anti-TPx-1 polyclonal antibodies. (D) Western blotting analysis of native TER ATPase using anti-TER ATPase polyclonal antibodies. (TIF)</p

Summary of the differentially abundant proteins in serum-derived exosomes from <i>E</i>. <i>multilocularis</i>-infected mice.

Summary of the differentially abundant proteins in serum-derived exosomes from E. multilocularis-infected mice.</p

The early diagnostic values of TPx-1 or TER ATPase in <i>E</i>. <i>multilocularis</i>-infected mice using ELISA.

For each antigen, the positive cut-off values were calculated by the OD value of each test serum (S) divided by the mean OD value of healthy sera (N). Samples with an S/N value not less than 2 were considered to be positive.</p