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Evaluation of the Efficiency of Detection of Bacterial DNA in Milk and Tissue Samples from Cattle, Sheep, and Goats by Conventional and Nested PCR Targeting COM1, SOD and Transposase IS1111 Genes of Coxiella burnetii Genome Cover

Evaluation of the Efficiency of Detection of Bacterial DNA in Milk and Tissue Samples from Cattle, Sheep, and Goats by Conventional and Nested PCR Targeting COM1, SOD and Transposase IS1111 Genes of Coxiella burnetii Genome

Open Access
|Apr 2025

Abstract

Q fever is a worldwide zoonosis, caused by Coxiella burnetii, an obligate intracellular bacterium that affects both humans and animals. The serious consequences on human health and the economic losses it causes, require the use of rapid, accurate, and sensitive diagnostic methods for its detection. PCR is the most widely used method for the molecular detection of Coxiella burnetii. Considering available information on the different sensitivity of PCR assays according to the selected genetic targets to be amplified, the present study aimed to compare the effectiveness of conventional and nested PCRs performed with primers Trans1/2, OMP1-4, and CB1/CB2 for the detection of Coxiella burnetii genome in samples, obtained from cattle, sheep and goats. Thirty archival DNAs, extracted from placentae, vaginal swabs, bulk tank milk samples, and cheese were tested. The highest level of detection was found when samples were tested with nested PCR with primers OMP1-4, targeting the Com1 gene (96.3%), and to a lesser extent with conventional PCR (56.7% positivity), performed with primers Trans1/2, encompassing a part of the IS1111 insertion sequence. A correlation was found between the detection efficiency of some primers and the type and origin of the samples. The results show that the sensitivity of the various PCR protocols for the detection of Coxiella burnetii could vary, thus the results obtained with one genetic marker should be interpreted with caution.

Language: English
Page range: 137 - 146
Submitted on: Jun 21, 2024
Accepted on: Feb 28, 2025
Published on: Apr 16, 2025
Published by: Ss. Cyril and Methodius University in Skopje
In partnership with: Paradigm Publishing Services
Publication frequency: 2 issues per year

© 2025 Konstantin Borisov Simeonov, Keytlin Venelinova Todorova, Petia Dinkova Genova-Kalou, published by Ss. Cyril and Methodius University in Skopje
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.