Vet World   Vol.15   July-2022  Article-23

Research Article

Veterinary World, 15(7): 1772-1778

https://doi.org/10.14202/vetworld.2022.1772-1778

Comparison of ITS-1 and TBR-1/2 primer sensitivity for the detection of Trypanosoma evansi local isolates in experimental rats using a polymerase chain reaction

Endang Suprihati1, Lucia Tri Suwanti1, Aditya Yudhana1,2, and Andika Indra Kusumaningrum3
1. Department of Veterinary Science, Division of Veterinary Parasitology, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, East Java, Indonesia.
2. Department of Veterinary Medicine, School of Health and Life Sciences, Universitas Airlangga, Banyuwangi, East Java, Indonesia.
3. Department of Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, East Java, Indonesia.

Background and Aim: Surra is caused by Trypanosoma evansi. The detection method using conventional parasitological tests has not always shown positive results in blood parasite detection, although the livestock has presented with clinical signs. Therefore, a fast and accurate diagnosis is necessary to prevent the disease predominately in field isolates. This study aimed to investigate the sensitivity of molecular detection method using two different specific primers, namely, Internal Transcribed Spacer 1 (ITS-1) and Trypanosoma brucei repeat 1/2 (TBR-1/2) against T. evansi field isolates from Banten Province, Indonesia.

Materials and Methods: The isolates of T. evansi used in this study were collected from Banten Province and cultured and preserved by the National Research Center for Veterinary Science, Indonesia. Eighteen experimental rats were divided into three equal groups, which were categorized as control, 1 × 101, and 1 × 104 infective doses. The isolates were injected into all experimental albino rats intraperitoneally. All samples were tested using conventional blood smear, card agglutination test (CATT), and polymerase chain reaction (PCR) method.

Results: The results of the CATT examination in all treatments showed negative results. However, PCR results showed that two different primers, namely, ITS-1 and TBR-1/2 had been successfully detected T. evansi from infected experimental rats, proven by positive PCR band appeared in 480 base pairs (bp) and 164 bp, respectively.

Conclusion: Based on the molecular diagnostic test using PCR method, TBR-1/2 primer is more sensitive to detect T. evansi compared to ITS-1 primer. The present finding provides preliminary data for studying the efficiency of different primers if practically applied as a standard diagnostic test for trypanosomiasis, especially in Indonesian livestock. Keywords: infectious disease, ITS-1, surra, TBR-1/2, tropical disease.

Keywords: infectious disease, ITS-1, surra, TBR-1/2, tropical disease.

How to cite this article: Suprihati E, Suwanti LT, Yudhana A, Kusumaningrum AI (2022) Comparison of ITS-1 and TBR-1/2 primer sensitivity for the detection of Trypanosoma evansi local isolates in experimental rats using a polymerase chain reaction, Veterinary World, 15(7): 1772–1778.

Received: 28-11-2021  Accepted: 17-06-2022     Published online: 24-07-2022

Corresponding author: Endang Suprihati   E-mail: esuprihati@yahoo.co.id

DOI: 10.14202/vetworld.2022.1772-1778

Copyright: Suprihati, 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.