Sığırlarda Babesia bovis ve Babesia bigemina’nın Reverse Line Blotting, Nested PCR ve Real Time PCR Teknikleri İle Karşılaştırmalı Tanısı

Abstract

SummaryThis study was carried out to compare Reverse Line Blotting (RLB), Nested PCR and Real Time PCR techniques in the molecular diagnosis of Babesia bovis and Babesia bigemina in cattle. Genomic DNA extractions were performed on 400 blood samples which were previously collected from cattle in various provinces of Turkey and stored in the laboratory with respect to use in different project studies. The concentrations of the DNAs were measured in NanoDrop spectrophotometer and analyzed by RLB, Nested PCR and Real Time PCR techniques after preparing the suitable concentrations. Totally 18 (4.50%), 59 (14.75%), 16 (4.00%) and 2 (0.50%) of examined samples were found to be infected with B. bovis, B. bigemina, Babesia spp. and B. bovis + B. bigemina mix, respectively by RLB. 23 (5.75%), 71 (17.75%), 7 (1.75%) and 23 (5.75%), 75 (18.75%), 9 (2.00%) of the examined samples were found to be infected with B. bovis, B. bigemina and B. bovis + B. bigemina mix by Nested PCR and Real Time PCR, respectively. When comparing the Nested PCR and RLB results with Real Time PCR assay, 94.4% and 88.8% sensitivity and both 100.0% specificity were determined, respectively. 5, 9 and 2 out of the total 16 Babesia spp. positivity's in RLB test were determined as B. bigemina, B. bovis, B. bovis + B. bigemina mix, respectively by both Real Time and Nested PCR. In conclusion, Real Time PCR was found to be more sensitive than Nested PCR and RLB in the investigation of B. bovis and B. bigemina in cattle with this study.

Sivas Yöresinde Sığır ve Kenelerde Theileria, Babesia ve Anaplasma Enfeksiyonlarının Araştırılması

“…and Babesia spp.) and 10 species specific probes (T. annulata, T. buffeli, B. bigemina, B. bovis, B. divergens, B. major, A. centrale, A. marginale, A. bovis and A. phagocytophilum) were used in RLB ( There are many molecular-based studies aimed at detecting the presence and prevalence of theilerioisis, babesiosis, anaplasmois in cattle and ixodid ticks from Turkey (Aktas et., 2011;Altay et al, 2008a;2014;Ica et al, 2007a;2007b;Yildirim et al, 2013). However, there is not any molecular-based research on tick-borne parasites in cattle in Sivas.…”

“…In addition, two new isolates, one of which was in Babesia genus and the other in Anaplasma genus, were found in the region. There are molecular-based studies showing the presence of new tick-borne parasite isolates in Turkey (Altay et al, 2008a;Ica et al, 2007b There are many molecular-based studies aimed at detecting the presence and prevalence of theilerioisis, babesiosis, anaplasmois in cattle and ixodid ticks from Turkey (Aktas et., 2011;Altay et al, 2008a;2014;Ica et al, 2007a;2007b;Yildirim et al, 2013). However, there is not any molecular-based research on tick-borne parasites in cattle in Sivas.…”

“…Molecular diagnostic methods such as polymerase chain reaction (PCR) and PCR-based reverse line blotting (RLB) have become widely used as sensitive and specific tools for detection and discrimination of tick-borne parasites in both their vectors and hosts (Aktas et al, 2006;Aktas and Ozubek, 2015;Altay et al, 2008aAltay et al, , 2008bAltay et al, , 2008cAltay et al, 2012;Altay et al,2014;Ica et al, 2007b;Yildirim et al, 2013). The capability of simultaneous detection of multiple pathogens in one sample is the superiority of reverse line blotting (Altay et al, 2008a) Babesiosis, theileriosis and anaplasmosis are the main tick-borne diseases in Turkey (Inci et al, 2016).…”

Bovine Babesiosis in Turkey: Impact, Current Gaps, and Opportunities for Intervention

“…After that, serological studies based on Indirect Fluorescent Antibody Technique (IFAT) revealed exposure to B. bovis , B. bigemina , and B. divergens parasites in different regions of Turkey [ 108 , 109 , 110 ]. Molecular techniques, such as PCR and RLB, have been recently used to investigate the presence of bovine babesiosis in the country [ 100 , 111 , 112 ]. Combined serology and molecular results have shown that bovine babesiosis caused by B. bovis , B. bigemina , B. divergens , B. major , and B. occultans is widespread in several areas of Turkey ( Table 2 and Figure 1 ).…”