Abstract
BackgroundDetection of Trypanosoma cruzi antigens in clinical samples is considered an important diagnostic tool for Chagas disease. The production and use of polyclonal antibodies may contribute to an increase in the sensitivity of immunodiagnosis of Chagas disease.Methodology/Principal findingsPolyclonal antibodies were raised in alpacas, rabbits, and hens immunized with trypomastigote excreted-secreted antigen, membrane proteins, trypomastigote lysate antigen and recombinant 1F8 to produce polyclonal antibodies. Western blot analysis was performed to determine specificity of the developed antibodies. An antigen capture ELISA of circulating antigens in serum, plasma and urine samples was developed using IgY polyclonal antibodies against T. cruzi membrane antigens (capture antibody) and IgG from alpaca raised against TESA. A total of 33 serum, 23 plasma and 9 urine samples were analyzed using the developed test. Among serum samples, compared to serology, the antigen capture ELISA tested positive in 55% of samples. All plasma samples from serology positive subjects were positive in the antigen capture ELISA. All urine positive samples had corresponding plasma samples that were also positive when tested by the antigen capture ELISA.ConclusionsPolyclonal antibodies are useful for detection of circulating antigens in both the plasma and urine of infected individuals. Detection of antigens is direct evidence of the presence of the parasite, and could be a better surrogate of current infection status.
Highlights
Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is endemic to many parts of the Americas [1,2,3]
This study describes the development of polyclonal antibodies raised against different Trypanosoma cruzi antigens and their applicability for the diagnosis of Chagas disease using the widely-used ELISA format
When trypomastigote lysate antigen (TLA) was used as antigen for western blot and tested with IgG3 from alpaca immunized with TLA, two proteins bands of 40 and 50 kDa were recognized
Summary
Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is endemic to many parts of the Americas [1,2,3]. This parasite infects a wide variety of wild and domestic mammals including humans [2]. The disease is transmitted by insect vectors (members of the Triatominae family) with metacyclic trypomastigotes present in their feces. It has been estimated that Chagas disease affects approximately 8 million people and may cause about 12,000 deaths each year (45,000 in the 1980s and 23,000 in the 1990s) [2,5]. Detection of Trypanosoma cruzi antigens in clinical samples is considered an important diagnostic tool for Chagas disease. The production and use of polyclonal antibodies may contribute to an increase in the sensitivity of immunodiagnosis of Chagas disease
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