Kamel Manal, Salah Faten, Demerdash Zeinab, Maher Sara, El-Baz Hanan, Zakaria Nahla, Abu-Shady Rania, Saad Amany, Hassan Salwa, Aziz Doaa Abdel
Immunology Department, Theodor Bilharz Research Institute, Kornish El Nil Street, Giza, Egypt.
Clinical Pathology, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
Acta Parasitol. 2024 Dec;69(4):1926-1936. doi: 10.1007/s11686-024-00917-9. Epub 2024 Oct 2.
For years, the Kato-Katz (KK) technique has been considered the gold standard for diagnosing schistosomiasis. The aim of this study was to compare the effectiveness of our previously developed gold nanoparticle-based lateral flow test strip (AuNPs-LFTS) for diagnosing active Schistosoma mansoni with that of the commercially available point-of-care Circulating Cathodic Antigen detection (POC-CCA) kit.
In this study, we collected sixty positive and twenty negative urine samples from patients in endemic hot spots in the Nile Delta, as well as from patients visiting the internal medicine clinic at Theodor Bilharz Research Institute (TBRI). We produced monoclonal antibodies (MAbs) against S. mansoni soluble egg antigen (SEA) from cloned hybridoma cells (4D/1D). These MAbs were conjugated with gold and mesoporous silica nanoparticles, and used to develop the LFTS.
The LFTS demonstrated a limit of detection (LoD) of 3 ng/ml. The sensitivity and specificity of the developed LFTS were found to be 96.7% and 95%, respectively, compared to 85% and 90% for the POC-CCA detection kit. The cases were divided into groups based on egg count in the stool, categorized as light, moderate, and heavy infections. The sensitivity of the LFTS in the group with light infection was higher than that of the POC-CCA. When using the KK technique (eggs per gram of stool sample [EPG]) as the reference test, the kappa value for the nano-based strips was 0.902, compared to 0.672 for the CCA strips, indicating an almost perfect agreement between KK and our developed LFTS.
These results confirm the reliability and effectiveness of the LFTS compared to commercially available kits for rapid, sensitive, and early diagnosis of schistosomiasis. However, it is recommended to conduct further assessments of the developed strip on a larger scale with a broader range of cases before considering its introduction to local or international markets.
多年来，加藤厚涂片法（KK法）一直被视为诊断血吸虫病的金标准。本研究旨在比较我们先前开发的基于金纳米颗粒的侧向流动试纸条（AuNPs-LFTS）与市售即时检测循环阴极抗原检测（POC-CCA）试剂盒在诊断曼氏血吸虫现症感染方面的有效性。
在本研究中，我们从尼罗河三角洲的流行热点地区患者以及 Theodor Bilharz 研究所（TBRI）内科门诊的患者中收集了60份阳性和20份阴性尿液样本。我们从克隆的杂交瘤细胞（4D/1D）中制备了针对曼氏血吸虫可溶性虫卵抗原（SEA）的单克隆抗体（MAbs）。这些单克隆抗体与金纳米颗粒和介孔二氧化硅纳米颗粒偶联，并用于开发试纸条。
该试纸条的检测限（LoD）为3 ng/ml。已开发的试纸条的灵敏度和特异性分别为96.7%和95%，而POC-CCA检测试剂盒的灵敏度和特异性分别为85%和90%。根据粪便中的虫卵计数将病例分为轻、中、重度感染组。试纸条在轻度感染组中的灵敏度高于POC-CCA。以KK法（每克粪便样本中的虫卵数[EPG]）作为参考检测方法时，纳米试纸条的kappa值为0.902，而CCA试纸条的kappa值为0.672，这表明KK法与我们开发的试纸条之间几乎完全一致。
这些结果证实了与市售试剂盒相比，试纸条在血吸虫病快速、灵敏和早期诊断方面的可靠性和有效性。然而，在考虑将其引入本地或国际市场之前，建议对开发的试纸条进行更大规模、更广泛病例的进一步评估。
