ArticleParasites & vectors2017
Improved molecular detection of Babesia infections in animals using a novel quantitative real-time PCR diagnostic assay targeting mitochondrial DNA.
Article in Parasites & vectors, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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41 citing papers in PubMed, 80 citations in OpenAlex.
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- A One-Pot RPA-CRISPR/Cas12a Assay for Rapid Genus-Level Detection of Babesia spp. in Ticks and Livestock Blood Samples.Transboundary and emerging diseases · 2026Article
- CRISPR-Cas12a-based rapid detection of Babesia gibsoni and Ehrlichia canis in dogs using fluorometer platform.Scientific reports · 2025Article
- Isolation and Characterization of Rickettsia finnyi, Novel Pathogenic Spotted Fever Group Rickettsia in Dogs, United States.Emerging infectious diseases · 2025Article
- Comparative molecular detection and phylogenetic analysis ofVeterinary world · 2025Article
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- Molecular Diagnosis and Identification of Equine Piroplasms: Challenges and Insights from a Study in Northern Italy.Animals : an open access journal from MDPI · 2025Article
- Comparative evaluation of real-time polymerase chain reaction, conventional PCR and microscopy for detection ofJournal of parasitic diseases : official organ of the Indian Society for Parasitology · 2024Article
- Contrasting pathogen prevalence between tick and dog populations at Chornobyl.Parasites & vectors · 2024Article
- A set of diagnostic tests for detection of active Babesia duncani infection.International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases · 2024Article
- Unveiling genotypic diversity of Theileria orientalis in lethal outbreaks among bovines in Karnataka, India.Parasitology research · 2024Article
- Germany-wide citizen science study reveals spread ofCurrent research in parasitology & vector-borne diseases · 2024Article
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- Molecular survey of potentially pathogenic microorganisms in ticks collected from coatis (Nasua nasua) in Iguaçu National Park, Atlantic Forest biome, southern Brazil.Parasitology research · 2023Article
- Canine piroplasmids: Molecular detection and laboratory characterization in dogs from Brasilia, Brazil, with the first molecular evidence of dog exposure to a novel opossum-associated Babesia sp.Ticks and tick-borne diseases · 2023Article
- Epidemiological analysis of cattle ticks and tick-borne pathogens in Gadag district, Karnataka state in India.Journal of parasitic diseases : official organ of the Indian Society for Parasitology · 2023Article
- Detection ofPathogens (Basel, Switzerland) · 2023Article
- A retrospective study of vector borne disease prevalence among anemic dogs in North Carolina.PloS one · 2023Article
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundBabesiosis is a protozoal, tick transmitted disease found worldwide in humans, wildlife and domesticated animals. Commonly used approaches to diagnose babesiosis include microscopic examination of peripheral blood smears, detection of circulating antibodies and PCR. To screen and differentiate canine Babesia infections many PCR assays amplify the 18S rRNA gene. These sequences contain hypervariable regions flanked by highly conserved regions allowing for amplification of a broad-range of Babesia spp. However, differences in the 18S rRNA gene sequence of distantly related clades can make it difficult to design assays that will amplify all Babesia species while excluding the amplification of other eukaryotes. By targeting Babesia mitochondrial genome (mtDNA), we designed a novel three primer qPCR with greater sensitivity and broader screening capabilities to diagnose and differentiate Babesia spp.
methodsUsing 13 Babesia mtDNA sequences, a region spanning two large subunit rRNA gene fragments (lsu5-lsu4) was aligned to design three primers for use in a qPCR assay (LSU qPCR) capable of amplifying a wide range of Babesia spp. Plasmid clones were generated and used as standards to determine efficiency, linear dynamic range and analytical sensitivity. Animals naturally infected with vector-borne pathogens were tested retrospectively and prospectively to determine relative clinical sensitivity and specificity by comparing the LSU qPCR to an established 18S rDNA qPCR.
resultsThe LSU qPCR efficiencies ranged between 92 and 100% with the limit of detection at five copies/reaction. The assay did not amplify mammalian host or other vector-borne pathogen gDNA except Cytauxzoon felis (a feline protozoal pathogen). The LSU qPCR assay amplified 12 different Babesia. sp. and C. felis from 31/31 (100%) archived samples, whereas the 18S qPCR amplified only 26/31 (83.9%). By prospective analysis, 19/394 diagnostic accessions (4.8%) were LSU qPCR positive, compared to 11/394 (2.8%) 18S rDNA qPCR positive.
conclusionsWe have developed a more sensitive qPCR assay with a more expansive range of Babesia spp. detection by targeting a highly conserved region of mtDNA, when compared to an established 18S qPCR.
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