Molecular Characterization of Echinococcus granulosus isolates from Human Cases Using Gold Nanoparticles-Based DNA Microarray with Silver Enhancement Simple Colorimetric Technology

Document Type : Original Article

Authors

1 Department of Medical Parasitology, Faculty of Medicine, Cairo University, Egypt

2 Department of Medical Parasitology, Faculty of Medicine, Cairo University, Egypt -Department of Endemic Medicine, Armed Forces Collage of Medicine (AFCM), Cairo

3 Department of Endemic Medicine, Faculty of Medicine, Cairo University, Egypt

4 Department of HBP Surgery, National Hepatology and Tropical Medicine Research Institute (NHTMRI), Cairo, Egypt

5 Department of Clinical Pathology, Faculty of Medicine, Cairo University, Egypt

6 Molecular Genetics and Molecular Diagnosis, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Egypt

7 Department of Molecular Biology, Medical Research Centre, Faculty of Medicine, Ain Shams University, Cairo, Egypt

Abstract

The Middle East including Egypt is considered as an endemic area for hydatid disease which predominantly affects the liver. Identification of Echinococcus strains is essential for proper vaccines development and control programs. The current study described the development of gold nanoparticles (AuNP) based genomic microarray that specifically identifies specific COX1 mitochondrial gene and concurrently provides genetic typing of E. granulosus complex without amplification process, aided by colorimetric silver enhancement technology. Signals are initiated by silver precipitation on AuNP combined with streptavidin, utilized to detect the biotinylated DNA. The qualitative purpose of our technique enables a simple detection by CCD camera connected to a computer. Thus, proposing a relatively easy and cost-effective alternative to the complex DNA amplification system can be applied in low-equipped laboratories.  In this work, a total of 30 samples from human hydatid cysts were investigated by oligonucleotide microarray with 10 amino labelled probes, corresponding to the 10 commonly known E. granulosus genotypes to be subsequently confirmed by DNA sequencing. Accordingly, camel strain, G6 was the predominant source of human hydatidosis among our cases and was found in 19 cases out of 30 (63.3%), while G1 genotype was reported among the remaining cases (36.7%). There was a statistically significant association between G6 and positive serological results plus multiple organs’ involvement, possibly indicating its superior antigenic effect. The predominance of the camel strain in Egyptian hydatidosis highlights the impact of ecological factors on camels’ importation strategies with the recommendation of considering vaccination to the imported animals to break the parasite cycle.

Keywords