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Development and Evaluation of a Semi-Nested PCR Method Based on the 18S ribosomal RNA Gene for the Detection of Babesia aktasi Infections in Goats

Authors :
Mehmet Can Ulucesme
Sezayi Ozubek
Munir Aktas
Source :
Veterinary Sciences, Vol 11, Iss 10, p 466 (2024)
Publication Year :
2024
Publisher :
MDPI AG, 2024.

Abstract

We developed and evaluated a semi-nested PCR assay for the detection of Babesia aktasi infection in goats based on the sequence of the B. aktasi 18S ribosomal RNA gene. Following in silico screening, the specificity of the primers was assessed using reference DNA samples, including B. ovis, B. motasi, B. crassa, B. venatorum, B. divergens, B. capreoli, Theileria ovis, and T. annulata. To determine the sensitivity of the method, blood infected with 2% parasitemia of B. aktasi was diluted to 10-fold serial dilutions. The method specifically amplified a 438 bp fragment of B. aktasi DNA, but did not demonstrate cross-amplification with the other hemoparasites tested. The sensitivity assay indicated that this PCR method was able to detect infection at a dilution of 10−8 of 2% parasitemia (0.074 parasites/200 µL). Ninety-seven blood samples collected from goats were used to analyze for B. aktasi, and the infection was detected in 18.5% of the goats. Additionally, the method was also applied to 44 field DNA samples that were detected to be positive for B. aktasi by reverse line blotting (RLB), and showed 84.1% agreement. The findings revealed that newly developed semi-nested PCR can detect B. aktasi infections in goats with high sensitivity and specificity.

Details

Language :
English
ISSN :
23067381
Volume :
11
Issue :
10
Database :
Directory of Open Access Journals
Journal :
Veterinary Sciences
Publication Type :
Academic Journal
Accession number :
edsdoj.143ca12ca58c4258b58b7b13efbed2a7
Document Type :
article
Full Text :
https://doi.org/10.3390/vetsci11100466