Abstract
The aim of this study was to optimize a PCR assay that amplifies an 843 pb fragment from the p28 gene of Ehrlichia canis and compare it with two other PCR methods used to amplify portions of the 16S rRNA and dsb genes of Ehrlichia. Blood samples were collected from dogs suspected of having a positive diagnosis for canine ehrlichiosis. Amplification of the p28 gene by PCR produced an 843-bp fragment and this assay could detect DNA from one gene copy among 1 billion cells. All positive samples detected by the p28-based PCR were also positive by the 16S rRNA nested-PCR and also by the dsb-based PCR. Among the p28-based PCR negative samples, 55.3% were co-negatives, but 27.6% were positive in 16S rRNA and dsb based PCR assays. The p28-based PCR seems to be a useful test for the molecular detection of E. canis, however improvements in this PCR sensitivity are desired, so that it can become an important alternative in the diagnosis of canine ehrlichiosis.
Highlights
Canine monocitic ehrlichiosis is an important tick-borne disease caused by the obligate intracellular bacterium Ehrlichia canis, which has worldwide distribution especially in tropical and subtropical areas
The purpose of this study was to develop a PCR assay based on amplification of the E. canis p28 gene and to compare this technique with two PCR methods used to amplify the 16S rRNA and dsb genes of Ehrlichia
Detection of E. canis p28 gene by PCR was optimized with genomic DNA extracted from the blood sample of an infected dog
Summary
Canine monocitic ehrlichiosis is an important tick-borne disease caused by the obligate intracellular bacterium Ehrlichia canis, which has worldwide distribution especially in tropical and subtropical areas. Because of the disadvantages of these methods, PCR has been used as a complement in the diagnosis of canine ehrlichiosis, due to its high sensitivity and specificity for detection of low levels of E. canis DNA (Iqbal; Rikihisa, 1994). This technique can be used to confirm active infection, and establish failure or success of treatment of ehrlichial infections (IQBAL; CHAICHANASIRIWITHAYA; RIKIHISA, 1994)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
