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Detecting the Lyme Disease Spirochete, Borrelia Burgdorferi, in Ticks Using Nested PCR
Published on: February 4, 2018
Differential detection of tick-borne pathogens in human platelets and whole blood using microfluidic PCR
Pavle Banović1, Elianne Piloto-Sardiñas2, Dragana Mijatović3
1Ambulance for Lyme Borreliosis and Other Tick-Borne Diseases, Department of Prevention of Rabies and Other Infectious Diseases, Pasteur Institute Novi Sad, Novi Sad, Serbia; Department of Microbiology With Parasitology and Immunology, Faculty of Medicine in Novi Sad, University of Novi Sad, Novi Sad, Serbia.
Abstract:
The tick-borne pathogens (TBPs) with adhesive phenotype can use platelets for dissemination and colonization of distant tissues and organs, and it has been shown that they can be found concentrated in the platelet fraction of blood. This study shows the differential presence of TBPs in samples of human platelet fraction (n = 68), whole blood samples (n = 68) and ticks collected (n = 76) from the same individuals, using an unbiased high-throughput pathogen detection microfluidic system. The clinical symptoms were characterized in enrolled patients. In patients with suspected TBP infection, serological assays were conducted to test for the presence of antibodies against specific TBPs. Tick species infesting humans were identified as Ixodes ricinus, Dermacentor reticulatus, and Haemaphysalis punctata. Eight patients developed local skin lesions at the site of the tick bite including non-specific lesions, itching sensation at the lesion site, and eschar. Most common TBPs detected in platelet fraction were Borrelia spielmanii and Rickettsia sp., followed by Borrelia afzelii and Anaplasma phagocytophilum. Multiple infections with three TBPs were detected in platelet fraction. In whole blood, most common TBPs detected were Anaplasma spp. and A. phagocytophilum, followed by Rickettsia spp. and B. afzelii. In ticks, the most common TBP detected was Rickettsia spp., followed by Borrelia spp. and Anaplasma spp. Overall, nine different pathogens with variable prevalence were identified using species-specific primers, and the most common was Rickettsia helvetica. In three patients, there were no coincidences between the TBPs detected in whole blood and tick samples. Only in one patient was detected A. phagocytophilum in both, whole blood and tick samples. These results suggest the unequal detection of TBPs in whole blood, platelet fraction and ticks collected, from the same individual. The results justify the use of both whole blood and platelet fraction for molecular diagnosis of TBPs in patients.
Insights
Tick-borne pathogens (TBPs) show varied presence in platelet fraction, whole blood, and ticks. Detecting TBPs in both whole blood and platelet fractions is crucial for accurate diagnosis.
Area of Science:
- Medical Microbiology
- Infectious Diseases
- Tick-borne Diseases
Background:
- Tick-borne pathogens (TBPs) can disseminate using platelets.
- Previous studies indicate TBPs concentrate in the platelet fraction of blood.
Purpose of the Study:
- To investigate the differential presence of TBPs in human platelet fraction, whole blood, and ticks from the same individuals.
- To correlate TBP detection with clinical symptoms and serological findings.
Main Methods:
- Utilized an unbiased high-throughput microfluidic system for pathogen detection.
- Analyzed samples from 68 patients (platelet fraction and whole blood) and 76 ticks.
- Identified tick species and conducted serological assays for antibodies against specific TBPs.
Main Results:
- Most common TBPs in platelet fraction: Borrelia spielmanii and Rickettsia sp. Multiple infections were detected.
- Anaplasma spp. and A. phagocytophilum were most common in whole blood.
- Rickettsia spp. was most prevalent in ticks; Rickettsia helvetica was the overall most common TBP detected.
- Significant discrepancies in TBP detection were observed across sample types.
Conclusions:
- Unequal detection of TBPs in whole blood, platelet fraction, and ticks highlights the complexity of TBP distribution.
- The findings support the combined use of whole blood and platelet fraction for optimal molecular diagnosis of TBPs.
