Development of Rapid Home-Made Immunochromatographic Test for Diagnosis of Cryptosporidium Parvum

Document Type : New and original researches in the field of Microbiology.

Authors

Department of Parasitology, Theodor Belharz Research Institute, Giza, Egypt

Abstract

Background: Immuno-chromatographic Test (ICT) devices have emerged as valuable tools for diagnosing parasitic infections. These tests are cost-effective, user-friendly, and require minimal training, making them ideal for use in settings with limited resources. Additionally, ICT offers rapid results with high accuracy, which makes it an attractive alternative to more complex diagnostic methods. Objective: This study aimed to develop a homemade immune-chromatographic kit utilizing nanotechnology for detecting specific antigens of Cryptosporidium parvum in human stool samples. Methodology: A comparative analysis was conducted between two diagnostic methods (sandwich ELISA and ICT) using 56 human stool samples confirmed to be infected with Cryptosporidium by Ziehl–Neelsen staining. The study also included 30 stool samples infected with other parasites and 20 healthy stool samples as negative controls. Silica nanoparticles were conjugated with anti-Cryptosporidium antibodies for enhanced sensitivity in both assays. Results: For sandwich ELISA, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 92.9%, 91%, 91.8%, and 92.6%, respectively, for the Cryptosporidium-infected group (52 out of 56 positive). Five healthy controls and other non-Cryptosporidium parasitic infections were falsely positive. In contrast, the homemade ICT detected 54 out of 56 Cryptosporidium-infected samples as positive, with two false positives from the control group. Conclusion: The ICT demonstrated superior sensitivity and specificity compared to sandwich ELISA, making it a more reliable, cost-effective, and accurate tool for diagnosing Cryptosporidium infections, especially in resource-limited settings.

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