A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for Anaplasma/Ehrlichia in Roaming Dogs and Symptomatic Dogs in Trinidad

This study compared two methods to detect cases of canine ehrlichiosis in a field setting. One method was a polymerase chain reaction for the 16S rRNA gene followed by reverse line blot hybridisation with genera and species-specific probes for Anaplasma/Ehrlichia. The second method was an autologous cell culture of peripheral leucocytes isolated from heparinised blood and maintained in a homologous canine serum in Dulbecco’s Modified Eagle medium without antibiotics. The cultures were examined under light microscopy for inclusion bodies after 48 h. Leucocytes were successfully propagated for 20 of the 34 samples submitted for autologous cell culture. Inclusion bodies were observed after cell culture in leucocytes of eight dogs. Two dogs were positive to the Anaplasma/Ehrlichia genera probe and six dogs were positive to the E. canis probe after reverse line blot hybridisation. There was acceptable agreement between reverse line blot hybridisation and cell culture results. Both reverse line blot hybridisation and autologous cell cultures can be used to detect E. canis in subclinical and clinical cases of disease. A definitive diagnosis of E. canis is best achieved by a combination of clinical signs, positive autologous cell culture, and reverse line blot hybridisation results.

Clinical comparison of two human papillomavirus detection assays: GenoFlow and reverse line blot

Introduction: Human papillomavirus (HPV) infection is typically critical in the oncogenesis of cervical cancer. However, available HPV detection kits differ in their ability and sensitivity to detect various types of HPV, and this variability has led to inconsistencies in the reporting of the geographic prevalence of HPV types, especially in developing countries. Here, we compared results of the recently developed GenoFlow HPV array test, which detects 33 HPV genotypes, to those of the well-established reverse line blot (RLB) assay, which detects 23 HPV types. Methodology: In total, 608 cervical specimens with cytology results ranging from normal to cancer were collected using an endocervical brush from women attending outpatient clinics in Riyadh, Saudi Arabia. Results: Sixty-nine specimens (11%) were positive for HPV. HPV genotype detection using the GenoFlow test had a sensitivity of 62% and a specificity of 100%. Overall agreement between the two HPV genotyping methods was 97%, with a concordance rate of 95%. Among the GenoFlow test results, 2% indicated additional HPV types that were not detected in the RLB assay, whereas the GenoFlow test missed 0.3% of the HPV types that were detected by the RLB; however, both tests were in agreement in detecting all major HPV types. Conclusion: The GenoFlow test was reliable, with results comparable to the RLB test. However, because the GenoFlow test is less labor-intensive and takes less total time (3 hours), it is a promising, affordable alternative to the RLB for HPV diagnosis and screening programs.

A Retrospective Epidemiological Study: The Prevalence of Ehrlichia canis and Babesia vogeli in Dogs in the Aegean Region of Turkey

Among tick-borne diseases, Ehrlichia canis and Babesia piroplasm cause important diseases in dogs where the distributions of the pathogen, vector and host overlap. The primary aim of the present study was to detect the prevalence of Babesia spp. and E. canis using PCR and reverse line blot (RLB) hybridization assay in a total of 379 samples comprising stray and owned dogs and to compare the diagnostic sensitivity of the two tests. Overall, 41.4% of dogs were infected with B. vogeli and/or E. canis as single (35.4%) and mixed (6.1%) infections. The majority of Babesia positive dogs (74.1%) were co-infected with E. canis. PCR detected a higher (P= 0.000) number of positivity in some provinces compared to RLB. To the best of our knowledge, these findings provide the first molecular evidence for the existence of B. vogeli in the Aegean Region, Turkey. The present study pinpoints the distribution and prevalence of E. canis and B. vogeli in the Aegean region of Turkey as of 2004 and as such establishes a baseline. This is of pivotal importance for future studies aimed to demonstrate changes in the dynamics of E. canis and B. vogeli infections in the region.