Iranian Veterinary Journal

Iranian Veterinary Journal

Innovative semi-quantitative multiplex single primer pair PCR method for simultaneous detecting some obligate intracellular piroplasms

Document Type : Research Paper

Authors
Zahra Alizadeh Sarabi 1
Nabian Nabian 2
Parviz Shayan 2
Saeid Bokaie 3
Hamid Reza Shokrani 4
1 PhD Student of Parasitology, Research Center for Ticks and Tick-borne Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
2 Professor, Department of Parasitology, Research Center for Ticks and Tick-borne Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
3 Professor, Departeman of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
4 Associate Professor, Department of Pathobiology, Lorestan University, Khorramabad, Iran
10.22055/ivj.2026.566326.2875
Abstract
The causative agents of piroplasmosis, Babesia spp. and Theileria spp. infect ruminants in the livestock industry with high economical loses worldwide. The early diagnosis of the disease can help manage the successful treatment. Giemsa staining is a common method used worldwide for diagnosis of piroplasmosis which unfortunately can be accompanied with some serious problems like suitable blood smears and quality of staining and needs high experience. Serological methods like ELISA or IFAT was also used in some laboratories. The molecular methods based on DNA analysis preferably used in many laboratories are accurate and reliable. The aim of the present study was to introduce a simple and innovative semi-quantitative multiplex PCR method which can be helpful for simultaneously detection of Babesia spp. and Theileria spp. For this aim, we used primers derived from 18S rRNA genes which can amplify simultaneously the DNA from hosts (ruminants and ticks) and also from Babesia spp., and Theileria spp with different easily distinguishable PCR products. Our results showed that, as was expected, the different length of ampilicons generated from Theileria spp. and Babesia spp. make it possible to differentiate these parasites from each other on the same agarose gel. Interestingly, since the same primer pair can also amplify the 18S rRNA from host cells with larger amplicon compared to the parasites, this property can be used for semi-quantitative analysis of the parasite burden in the samples. It is necessary to mention that the introduced primer pair (VetUTPiro) can be used for differential diagnosis of babesiosis and theileriosis as well as for semi-quantitative multiplex PCR analysis of the parasite burden consequently for controlled therapy management.
Keywords
Theileria spp
Babesia spp
Semi-quantitative multiplex PCR
Therapy management
Sheep
Subjects
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