IVD of Trypanosoma cruzi

Background

Chagas' disease is a parasitic disease caused by Trypanosoma cruzi and is transmitted by triatomine bugs and kissing bugs. In addition, Trypanosoma cruzi can be transmitted from mother to child or through contaminated needles. Trypanosoma cruzi is a fecal-borne trypanosome that invades the human body through broken skin, mucous membranes, or conjunctiva. Patients develop symptoms such as fever, lymphadenitis, and anemia after infection. Currently, Chagas' disease has become a serious public health problem that requires timely diagnosis and prevention.

ROS and RNS are produced throughout the life cycle of Trypanosoma cruziFig1. ROS and RNS are produced throughout the life cycle of Trypanosoma cruzi. (Mesías A C, et al., 2019)

Main Steps of IVD for Trypanosoma cruzi

  • Microscopic examination. Thick and thin blood smears were prepared and stained with Giemsa to observe trypanosomes.
  • Isolation and identification of pathogens. The mice can be inoculated with blood or cultured in a culture medium, and the growth of trypanosomes can be observed subsequently.
  • Immunological tests. Including indirect fluorescent antibody technology (IFA), indirect hemagglutination test (IHA), immunochromatography test strip method, and enzyme-linked immunosorbent assays (ELISA), which have high sensitivity.
  • Molecular biology technology. Use DNA probes and PCR technology to detect the variable region and small foreign genes of Trypanosoma cruzi small circle kDNA to obtain diagnostic results.
  • Peripheral blood picture. In the acute phase of infection, leukocytosis and erythrocyte sedimentation rate are accelerated.

Symptoms of Chagas' Disease

Acute Phase:

Often asymptomatic or presenting mild symptoms. Symptoms, when present, can include:

  • Fever
  • Swelling at the infection site (chagoma)
  • Rash
  • Anemia
  • Swelling of one eyelid (Romaña's sign if the parasite entered via the eye)
  • Lymphadenopathy (swollen lymph nodes)
  • Hepatosplenomegaly (enlarged liver and spleen)

Chronic Phase:

May remain asymptomatic for years or indefinitely in some people (indeterminate form). Chronic symptomatic form can affect various organs, notably the heart and digestive system, leading to:

  • Cardiomyopathy (including heart failure, arrhythmias, and sudden death)
  • Megavisceras, like megacolon and megaesophagus, which can cause severe digestive issues.

Chagas' Disease Related Proteins

  • Cruzipain: A cysteine protease involved in the parasite's ability to degrade host proteins and evade the immune system.
  • Trans-sialidase: Enzymes that transfer sialic acids from host cell glycoconjugates to the parasite's surface, aiding in immune evasion.
  • Gp85/TS family: A family of glycoproteins involved in host cell adhesion and invasion.
  • TcISP2 and TcISP3: Inhibitors of serine peptidases, which play a role in modulating host immune responses.
  • TcTASV (Trypomastigote Surface Antigen Variant): Comprising several proteins and related to the surface antigen variants of trypomastigotes.
  • TcGP63: A glycoprotein utilized by T. cruzi for proteolytic degradation and immune modulation.

Creative BioMart provides high-quality recombinant Trypanosoma cruzi related proteins used for IVD, including ELISA, lateral flow assay, western blot, and other immunoassays.

Highlights of Our Products

  • High sensitivity and specificity. The protein can accurately identify and detect specific biomolecules or disease markers.
  • Highly repeatable.
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  • It is widely used and suitable for downstream immunological experiments.
  • Outstanding success rate and fast development speed.

Our Outstanding Advantages

  • We are committed to offering IVD proteins with high performance, reliability, and consistency that exceed industry-leading levels.
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Applications of Chagas' Disease Related Proteins

Diagnostic Kits and Biomarkers: Proteins like cruzipain, trans-sialidase, and various surface antigens are key targets for the development of diagnostic tests. ELISA, rapid tests, and PCR can detect antibodies or antigens related to T. cruzi.

Vaccine Development: Identifying antigenic proteins such as those from the gp85/TS family can be crucial for developing vaccines. These proteins can elicit an immune response to provide protective immunity.

Drug Targets: Many Chagas' related proteins are considered potential drug targets. Cruzipain and trans-sialidase, for example, have been targeted for the development of inhibitors that could potentially treat T. cruzi infections.

Therapeutic Antibodies: Monoclonal antibodies against specific T. cruzi proteins can be developed for therapeutic purposes, potentially neutralizing the parasite or preventing its invasion and dissemination.

Research Tools: These proteins can be used to develop research tools, including genetic constructs, recombinant proteins, and immune reagents, to further study the biology and pathology of T. cruzi and Chagas' disease.

Case Study

Case 1: Pascual-Vázquez G, Alonso-Sardón M, Rodríguez-Alonso B, et al. Molecular diagnosis of Chagas disease: a systematic review and meta-analysis. Infect Dis Poverty. 2023 Oct 16;12(1):95. doi: 10.1186/s40249-023-01143-7. PMID: 37845734; PMCID: PMC10577976.

The authors identified 858 records and selected 32 papers. Studies pertained to endemic countries and nonendemic areas with adult and paediatric populations. The sample sizes ranged from 17 to 708 patients. There were no concerns regarding the risk of bias and applicability of all included studies. This resulted in high heterogeneity between studies, with the master mix origin and guanidine addition representing significant sources.

Fig2. Graph of the number of papers included in this systematic review

Case 2: Ferreira AZL, de Araújo CN, Cardoso ICC, de Souza Mangabeira KS, Rocha AP, Charneau S, Santana JM, Motta FN, Bastos IMD. Metacyclogenesis as the Starting Point of Chagas Disease. Int J Mol Sci. 2023 Dec 21;25(1):117. doi: 10.3390/ijms25010117. PMID: 38203289; PMCID: PMC10778605.

This review provides an overview of the current knowledge regarding the parasite's life cycle, molecular pathways, and mechanisms involved in metabolic and morphological adaptations during MTG, enabling the MT to evade the immune system and successfully infect human cells.

Fig3. idirectional signalling pathways activated in both MT and the host cell during invasion. Trans-sialidases transfer sialic acid from the host cell membrane to mucins or mucin-like glycoproteins on the surface of the parasite, a process necessary for parasite adhesion and penetration. GP82 binds to its receptor LAMP2 and triggers cleavage of PIP2 through the activation of PLC, generating IP3 and DAG, leading to Ca2+ release to the cytosol and PKC translocation to the membrane. Phosphorylated PKC activates ERK1/2 and mTOR dephosphorylation, leading to TFEB translocation, actin disruption, and lysosome recruitment. Treatment with MVK negatively modulates invasion by inhibiting parasite internalisation through the phosphorylation and activation of MAPK. Extracellular SAPs bind to LAMP2 and activate cytosolic Ca2+ mobilisation. Ultimately, this will result in the formation of the parasitophorous vacuole and host cell invasion. GP90 acts as a downregulator of invasion and binds to the host cell without triggering a Ca2+ transient.

Case 3: Nunes JPS, Roda VMP, Andrieux P, Kalil J, Chevillard C, Cunha-Neto E. Inflammation and mitochondria in the pathogenesis of chronic Chagas disease cardiomyopathy. Exp Biol Med (Maywood). 2023 Nov;248(22):2062-2071. doi: 10.1177/15353702231220658. PMID: 38235691; PMCID: PMC10800136.

The authors summarized studies showing how IFN-γ and TNF-α affect cell energy generation, mitochondrial health, and redox homeostasis in cardiomyocytes, in addition to human CD and mitochondria. They hypothesize that cytokine-induced mitochondrial dysfunction in genetically predisposed patients may be the underlying cause of CCC severity and they believe this mechanism may have a bearing on other inflammatory cardiomyopathies.

Fig4. IFN-γ and TNF-α dysregulate energy production in mitochondria. Upon binding to their receptors, IFN-γ and TNF-α trigger the production of pro-inflammatory cytokines, including TNF-α and IL1-β, as well as the generation of nitric oxide (NO) by NOS2. This process occurs through the activation of the STAT1/NF-κB and MEK/MAPK/NF-κB pathways, respectively.