Main Article Content
Cystic echinococcosis is a zoonotic, neglected tropical disease caused by various genotypes of cestode parasite Echinococcus granulosus. Although, echinococcosis causes enormous financial and health impacts, no gold standard diagnostic technique is available. Genetic characterization of these prevalent cestode at species level is essential for disease management and appropriate control measures.
We aimed to investigate the genetic diversity of echinococcus granulosus using a modified Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) based assay in infected population.
A total of 18 human hydatid cyst samples were collected from various hospitals of Southern Punjab and Islamabad Capital Territory region of Pakistan. Extracted DNA was used for PCR amplification of mitochondrial NADH dehydrogenase subunit 1 (Nad1) gene followed by sequencing and phylogenetic analysis using Molecular evolutionary Genetic Analysis (MEGA) Software. The entire sequences were fed into NEBcutter V2.0 to select a single restriction enzyme followed by invitro confirmation through PCR-RFLP.
Amplification on the Nad1 gene was observed in 100% of the samples processed. The Basic Local Alignment Tool (BLAST) and phylogenetic tree analysis revealed 83.3% E. granulosus (G1-G3 genotypes), 11.1% E. multilocularis and 5.6% E. Canadensis (G6 genotype). The use of the BfaI enzyme in PCR-RFLP analysis revealed that all of the 18 samples were assigned consecutive genotypes as observed in the sequencing.
The current study concluded that the BfaI enzyme could be used for the genotypic analysis of echinococcosis in developing and frequently affected countries. It will be a cost-effective and easy technique compared to sequencing, which will aid in developing novel therapeutic and control strategies for the parasite.
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
All articles published in the Journal of Medical Sciences (JMS) are licensed under the Creative Commons Attribution 4.0 International License (CC-BY 4.0). Under the CC BY 4.0 license, author(s) retain the ownership of the copyright publishing rights without restrictions for their content, and allow others to copy, use, print, share, modify, and distribute the content of the article even for commercial purposes as long as the original authors and the journal are properly cited. No permission is required from the author/s or the publishers for this purpose. Appropriate attribution can be provided by simply citing the original article. The corresponding author has the right to grant on behalf of all authors, a worldwide license to JMS and its licensees in all forms, formats, and media (whether known now or created in the future), The corresponding author must certify and warrant the authorship and proprietorship and should declare that he/she has not granted or assigned any of the article’s rights to any other person or body.
The corresponding author must compensate the journal for any costs, expenses, or damages that the JMS may incur as a result of any breach of these warranties including any intentional or unintentional errors, omissions, copyright issues, or plagiarism. The editorial office must be notified upon submission if an article contains materials like text, pictures, tables, or graphs from other copyrighted sources. The JMS reserves the right to remove any images, figures, tables, or other content, from any article, whether before or after publication, if concerns are raised about copyright, license, or permissions and the authors are unable to provide documentation confirming that appropriate permissions were obtained for publication of the content in question.