Maleki Fatemeh, Akhlaghi Lame, Tabatabaie Fatemeh
School of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
Department of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Med J Islam Repub Iran. 2023 Aug 14;37:87. doi: 10.47176/mjiri.37.87. eCollection 2023.
Hydatidosis, a chronic zoonotic disease, has a distribution worldwide and is caused by the larval stage of the Echinococcus helminth. The Dot-ELISA test can diagnose hydatidosis quickly and accurately. Additionally, unlike other hydatid disease tests now used, this quick and affordable enzyme immunoassay is very serum-conservative and antigen-conservative, needing just nanogram levels of parasite antigen.
In the present cross-sectional study, crude and B antigens of hydatid cyst fluid were obtained to diagnose human hydatidosis using CIEP (Counter Immunoelectrophoresis), ELISA (Enzyme-linked Immuno Sorbent assay), and Dot- ELISA (Dot Enzyme linked Immuno Sorbent Assay) methods. Infected liver with a hydatid cyst was collected from Tehran's slaughterhouses to prepare cyst fluid in different stages. After extracting and purifying the Cyst fluid, it is centrifuged at 4ºc, then prepared to concentrate. The study also included sera from hydatidosis (n=60), samples of helminth parasites (n=55), fascioliasis (n=35), toxocariasis (n=20) and negative control (n=35) were tested by CIEP (Counter Immunoelectrophoresis), ELISA (Enzyme-linked Immune Sorbent assay), and Dot- ELISA (Dot Enzyme linked Immuno Sorbent Assay) methods. All statistical analyses were performed using Statistical Package for the Social Sciences (SPSS) for Windows release 25.0 (SPSS Inc., Chicago, IL, USA).
Crude antigen of hydatid cyst showed a specificity of 76.7%, a sensitivity of 93.3% using the ELISA method, and B antigen showed a specificity of 96.7% and sensitivity of 88.3% using the same method. The crude antigen of the hydatid cyst exhibited a specificity of 68.9% and a sensitivity of 86.7% using CIEP. The B antigen showed a specificity of 87.8% and sensitivity of 83.3% using the same method.The crude antigen of hydatid cyst having serum dilution at 1:800 exhibited a specificity of 83.3% and sensitivity of 100% using the Dot-ELISA method and B antigen having serum dilution at 1:800 serum showed a specificity of 100% and sensitivity of 98.3% using the same method. The results of this finding showed that B antigen has the maximum specificity to diagnose hydatid test using the Dot- ELISA method.
Hydatid cysts present with varied symptomatology. History of exposure to infected animals may not be present. A high degree of clinical suspicion combined with meticulous history and clinical examination supported by laboratory investigations are required for its diagnosis. The Dot-ELISA system with native antigen B is a viable approach for the immunodiagnosis of human hydatidosis that is preferred to infection.
包虫病是一种慢性人畜共患疾病,在全球范围内均有分布,由棘球绦虫的幼虫阶段引起。斑点酶联免疫吸附测定(Dot-ELISA)试验能够快速、准确地诊断包虫病。此外,与目前使用的其他包虫病检测方法不同,这种快速且经济实惠的酶免疫测定法对血清和抗原的需求量极少,仅需纳克级别的寄生虫抗原。
在本横断面研究中,采用对流免疫电泳(CIEP)、酶联免疫吸附测定(ELISA)和斑点酶联免疫吸附测定(Dot-ELISA)方法,获取包虫囊肿液的粗抗原和B抗原,以诊断人类包虫病。从德黑兰的屠宰场收集感染包虫囊肿的肝脏,制备不同阶段的囊肿液。提取并纯化囊肿液后,于4℃离心,然后进行浓缩制备。该研究还纳入了包虫病患者血清(n = 60)、蠕虫寄生虫样本(n = 55)、肝片吸虫病患者样本(n = 35)、弓蛔虫病患者样本(n = 20)以及阴性对照样本(n = 35),采用CIEP、ELISA和Dot-ELISA方法进行检测。所有统计分析均使用适用于Windows 25.0版本的社会科学统计软件包(SPSS)(SPSS公司,美国伊利诺伊州芝加哥)。
使用ELISA方法时,包虫囊肿的粗抗原特异性为76.7%,敏感性为93.3%;B抗原的特异性为96.7%,敏感性为88.3%。使用CIEP方法时,包虫囊肿的粗抗原特异性为68.9%,敏感性为86.7%;B抗原的特异性为87.8%,敏感性为83.3%。使用Dot-ELISA方法时,血清稀释度为1:800的包虫囊肿粗抗原特异性为83.3%,敏感性为100%;血清稀释度为1:800的B抗原特异性为100%,敏感性为98.3%。这一结果表明,使用Dot-ELISA方法诊断包虫病时,B抗原具有最高的特异性。
包虫囊肿的症状表现多样。患者可能没有接触过感染动物的病史。其诊断需要高度的临床怀疑,并结合细致的病史询问和临床检查,同时辅以实验室检查。采用天然抗原B的Dot-ELISA系统是一种可行的人类包虫病免疫诊断方法,优于感染诊断方法。