The Trypanosoma cruzi IgG ELISA is intended for the qualitative determination of IgG class antibodies against Chagas (Trypanosoma cruzi) in human serum or plasma (citrate, heparin). For Research Use Only-Not for Use in Diagnostic Procedures.
Contents of Kit
1. Reagents supplied a. Chagas (Trypanosoma cruzi) Coated Microplate (IgG): 12 break-apart 8-well snap-off strips coated with recombinant Chagas (Trypanosoma cruzi) antigens; in resealable aluminium foil. b. IgG Sample Diluent: 1 bottle containing 100 ml of phosphate buffer (10 mM) for sample dilution; pH 7.2 ± 0.2; coloured yellow; ready to use; white cap. c. Stop Solution: 1 bottle containing 15 ml sulphuric acid, 0.2 mol/l; ready to use; red cap. d. 20× Washing Buffer (20× conc.): 1 bottle containing 50 ml of a 20-fold concentrated phosphate buffer (0.2 M), pH 7.2 ± 0.2, for washing the wells; white cap. d. Protein A Conjugate: 1 bottle containing 20 ml of peroxidase labelled Protein A in phosphate buffer (10 mM); coloured blue; ready to use; black cap. e. TMB Substrate Solution: 1 bottle containing 15 ml 3,3',5,5'-tetramethylbenzidine (TMB), < 0.1 %; ready to use; yellow cap; < 5% NMP. f. Chagas (Trypanosoma cruzi) IgG Positive Control: 1 vial containing 2 ml control (human serum or plasma); coloured yellow; ready to use; red cap. g. Chagas (Trypanosoma cruzi) IgG Cut-off Control: 1 vial containing 3 ml control (human serum or plasma); coloured yellow; ready to use; green cap. h. Chagas (Trypanosoma cruzi) IgG Negative Control: 1 vial containing 2 ml control (human serum or plasma); coloured yellow; ready to use; blue cap. 2. Materials supplied a. 1 Cover foil b. 1 Instruction for use (IFU)
Storage
Store the kit at 2-8 °C. The opened reagents are stable up to the expiry date stated on the label when stored at 2-8 °C.
Precision
Citations
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Background
Chagas' disease is a prevalent infectious disease in Middle and South America transmitted to humans through the bite of infected reduviidae bugs. The disease is caused by a single-celled parasite called Trypanosoma cruzi. It can progress through different phases and often leads to a chronic condition. Approximately 4 to 5 million people in South America are affected, particularly those living in poor conditions. The disease can be fatal, with up to 10% of infections resulting in death, and newborns are at a higher risk.
Figure 1. Life cycle of Trypanosoma cruzi. (Source: Vazquez, M. P., 2007)
Transmission occurs when the parasite, present in the bug's feces, enters the human body through skin wounds. In utero transmission and blood transfusion are also possible routes of infection. Symptoms typically appear in 1-2% of infected individuals within one to four weeks, predominantly affecting children up to 15 years old. The acute phase is characterized by fever, diarrhea, abdominal pain, swollen lymph nodes, and generalized swelling. Neonates and infants may experience inflammation of the heart or brain. This phase lasts approximately four weeks and is more common in children. Following the acute phase, a latent period occurs with few or no symptoms, although immune system weakening may occur. This phase can last for several years. Around 10 to 20% of infected individuals progress to the chronic phase, which can affect various organs such as the heart, intestines, and nervous system. Chronic heart problems can lead to sudden cardiac death or heart failure, influencing the prognosis.
The Trypanosoma cruzi IgG ELISA kit is an important diagnostic tool for detecting past or present Chagas disease infection. This enzyme-linked immunosorbent assay (ELISA) is designed to qualitatively detect IgG antibodies against T. cruzi antigens in human serum or plasma samples. The detection is crucial for diagnosing Chagas' disease, particularly in the chronic phase where symptoms may be absent or non-specific. It helps identify individuals who require medical intervention, such as antiparasitic treatment, and enables monitoring of disease progression and treatment efficacy.
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References
Anti-Trypanosoma cruzi Immunoglobulin G1 can be a useful tool for diagnosis and prognosis of human Chagas' Disease
Clinical Diagnostic Laboratory Immunology
Authors: Cordeiro, F. D., Martins-Filho, O. A., Da Costa Rocha, M. O., Adad, S. J., Corrêa-Oliveira, R., Romanha, A. J.
Two functionally distinct antibodies, categorized as conventional serology antibodies (CSA) and lytic antibodies (LA) have been described in Chagas' disease, based on their ability to bind to fixed epimastigotes (EPI) or live trypomastigotes (TRYPO), respectively. In this study, the profile of immunoglobulin G (IgG) subclasses of CSA and LA were analyzed by flow cytometry using serum samples from chronic chagasic patients with the indeterminate (IND), cardiac (CARD), and digestive (DIG) clinical forms of the disease. The results were expressed as percentage of positive fluorescent parasites (PPFP) for each sample. CSA showed a higher PPFP than LA for all samples. At serum dilutions between 1:256 and 1:2,048, IgG1 anti-EPI was able to distinguish chagasic from nonchagasic individuals. Different profiles of IgG subclasses were observed for CSA and LA. IgG1 and IgG2 were the main subclasses in CSA, whereas IgG1 and IgG3 were the predominant ones in LA. The reactivity of IgG2 anti-EPI was greater in IND and CARD than in DIG patients. Furthermore, a low level of IgG1 and IgG3 LA was associated with most of the CARD patients. On the other hand, a high level of IgG1 LA was associated with most of the IND patients. In summary, our findings indicate the potential of IgG1 anti-EPI for serological diagnosis of Chagas' disease, providing further evidence for a protective role of LA, and show that IgG1 anti-liveTrypanosoma cruzi TRYPO may be used to predict the risk of cardiac damage in Chagas' disease.
In-house ELISA method to analyze anti-Trypanosoma cruzi IgG reactivity for differential diagnosis and evaluation of Chagas disease morbidity
Revista da Sociedade Brasileira de Medicina Tropical
Authors: Santos, L. D. S., Torres, R. M., Machado-de-Assis, G. F., Bahia, M. T., Martins, H. R., Teixeira-Carvalho, A., Lana, M. D.
The goal was to develop an in-house serological method with high specificity and sensitivity for diagnosis and monitoring of Chagas disease morbidity. METHODS: With this purpose, the reactivities of anti-T. cruzi IgG and subclasses were tested in successive serum dilutions of patients from Berilo municipality, Jequitinhonha Valley, Minas Gerais, Brazil. The performance of the in-house ELISA was also evaluated in samples from other relevant infectious diseases, including HIV, hepatitis C (HCV), syphilis (SYP), visceral leishmaniasis (VL), and American tegumentary leishmaniasis (ATL), and noninfected controls (NI). Further analysis was performed to evaluate the applicability of this in-house methodology for monitoring Chagas disease morbidity into three groups of patients: indeterminate (IND), cardiac (CARD), and digestive/mixed (DIG/Mix), based on their clinical status. RESULTS: The analysis of total IgG reactivity at serum dilution 1:40 was an excellent approach to Chagas disease diagnosis (100% sensitivity and specificity). The analysis of IgG subclasses showed cross-reactivity, mainly with NI, VL, and ATL, at all selected serum dilutions. Based on the data analysis, the IND group displayed higher IgG3 levels and the DIG/Mix group presented higher levels of total IgG as compared with the IND and CARD groups. CONCLUSIONS: These findings demonstrated that methodology presents promising applicability in the analysis of anti-T. cruzi IgG reactivity for the differential diagnosis and evaluation of Chagas disease morbidity.
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Figure 1. Life cycle of Trypanosoma cruzi.
