Development of a multiplex polymerase chain reaction technique for detection and discrimination of Eimeria spp. in cattle in Indonesia

Vet World Vol.15 April-2022 Article-22

Research Article

Veterinary World, 15(4): 975-980

https://doi.org/10.14202/vetworld.2022.975-980

Fitrine Ekawasti^1,2, Raden Wisnu Nurcahyo², Mukh Fajar Nashrulloh^2,3, Dwi Priyowidodo², and Joko Prastowo²

1. Indonesia Research Center for Veterinary Science, National Research and Innovation Agency, Bogor, 16114, Indonesia.
2. Department of Parasitology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia.
3. Applied Zoology Research Center, National Research and Innovation Agency, Bogor, 16911, Indonesia.

Background and Aim: Bovine eimeriosis is a disease caused by apicomplexan parasites of the genus Eimeria. It is one of the most important and widespread bovine illnesses in the world. Some of the identified species of bovine eimeriosis have morphologically similar oocysts that are difficult to differentiate. For the identification of particular Eimeria spp., diagnostic laboratories are increasingly turning to DNA-based technology. This study aims to develop a multiplex polymerase chain reaction (mPCR) technique based on the internal transcribed spacer-1 (ITS-1) gene for the simultaneous identification of pathogenic Eimeria spp. in cattle from Sulawesi Island, Indonesia.

Materials and Methods: Genomic DNA was extracted by the DNAzol reagent from the purified Eimeria oocysts. Species-specific primers targeting the ITS-1 region were used to amplify the distinct Eimeria spp.

Results: Using PCR ITS-1, this study showed that 36 of 120 fecal samples (30%) were infected by Eimeria spp. The multiplex PCR assay allowed for the simultaneous identification of six major Eimeria spp. in a single-tube reaction. The proportion of mixed Eimeria spp. infections was 100% (36/36). The maximum number of Eimeria spp. was five, and the minimum number was two.

Conclusion: Identification of six pathogenic Eimeria spp. in cattle was successfully carried out by nested multiplex PCR using ITS-1 gene. In the future, a procedure to detect pathogenic Eimeria spp. in one tube reaction will offer economical and save diagnostic time. Keywords: bovine, diagnostic, eimeriosis, multiplex, nested, Sulawesi island.

Keywords: bovine, diagnostic, eimeriosis, multiplex, nested, Sulawesi island.

How to cite this article: Ekawasti F, Nurcahyo RW, Nashrulloh MF, Priyowidodo D, Prastowo J (2022) Development of a multiplex polymerase chain reaction technique for detection and discrimination of Eimeria spp. in cattle in Indonesia, Veterinary World, 15(4): 975-980.

Received: 20-01-2022 Accepted: 10-03-2022 Published online: 18-04-2022

Corresponding author: Raden Wisnu Nurcahyo E-mail: wisnu-nc@ugm.ac.id

DOI: 10.14202/vetworld.2022.975-980

Copyright: Ekawasti, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.