Background

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.

Methodology/Principal findings

Polyclonal 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.

Conclusions

Polyclonal 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.

中文翻译：

---

用于检测克氏锥虫循环抗原的多克隆抗体。

背景

临床样品中克氏锥虫抗原的检测被认为是南美锥虫病的重要诊断工具。多克隆抗体的生产和使用可能有助于南美锥虫病免疫诊断的敏感性增加。

方法/主要发现

多聚体抗体在羊驼，兔和母鸡中产生，该母鸡用锥虫病的分泌分泌抗原，膜蛋白，锥虫病的溶胞产物抗原和重组1F8免疫，以产生多克隆抗体。进行蛋白质印迹分析以确定所开发抗体的特异性。使用针对T的IgY多克隆抗体，开发了血清，血浆和尿液样本中循环抗原的抗原捕获ELISA 。克鲁兹来自羊驼的膜抗原（捕获抗体）和IgG针对TESA。使用开发的测试分析了总共33个血清，23个血浆和9个尿液样本。在血清样本中，与血清学相比，抗原捕获ELISA在55％的样本中检测为阳性。来自血清学阳性受试者的所有血浆样品在抗原捕获ELISA中均为阳性。所有尿液阳性样品均具有相应的血浆样品，当通过抗原捕获ELISA测试时，血浆样品也为阳性。

结论

多克隆抗体可用于检测受感染个体的血浆和尿液中的循环抗原。抗原的检测是该寄生虫存在的直接证据，并且可以更好地替代当前的感染状况。