Development and evaluation of a PCR assay for the detection of Cytauxzoon felis DNA in feline blood samples

Birkenheuer, Adam J.; Marr, Henry S.; Alleman, A. Rick; Levy, Michael G.; Breitschwerdt, Edward B.

doi:10.1016/j.vetpar.2005.12.007

Cited by 47 publications

(41 citation statements)

References 17 publications

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“…Total RNA (10 µg/reaction) was treated twice with DNA- free DNase Treatment and Removal Reagent (Ambion, Grand Island, NY). Prior to generation of cDNA, the absence of contaminating DNA in the purified RNA was confirmed by PCR for C. felis 18S rRNA genes [3]. Cytauxzoon felis cDNA was produced using random hexamer primers (Promega, Madison WI) and Smartscribe reverse transcriptase (Clontech, Mountain View, CA).…”

Section: Methodsmentioning

confidence: 99%

“…There are currently no effective means to prevent cytauxzoonosis, and even with treatment costing thousands of dollars, up to 40% of cats still succumb [1], [2]. First described in Missouri in 1976, the geographic range of C. felis is expanding and it has now been diagnosed in domestic cats in one third of US states (Figure 1) [1], [3], [4], [5], [6], [7], [8], [9], [10], [11]. Expansion of the geographic range is presumed to be due to changes in climate, urbanization, and increased exposure to the bobcat [ Lynx rufus ] reservoir host and the tick vector [ Amblyomma americanum ].…”

Section: Introductionmentioning

confidence: 99%

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A Novel Candidate Vaccine for Cytauxzoonosis Inferred from Comparative Apicomplexan Genomics

et al. 2013

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Cytauxzoonosis is an emerging infectious disease of domestic cats (Felis catus) caused by the apicomplexan protozoan parasite Cytauxzoon felis. The growing epidemic, with its high morbidity and mortality points to the need for a protective vaccine against cytauxzoonosis. Unfortunately, the causative agent has yet to be cultured continuously in vitro, rendering traditional vaccine development approaches beyond reach. Here we report the use of comparative genomics to computationally and experimentally interpret the C. felis genome to identify a novel candidate vaccine antigen for cytauxzoonosis. As a starting point we sequenced, assembled, and annotated the C. felis genome and the proteins it encodes. Whole genome alignment revealed considerable conserved synteny with other apicomplexans. In particular, alignments with the bovine parasite Theileria parva revealed that a C. felis gene, cf76, is syntenic to p67 (the leading vaccine candidate for bovine theileriosis), despite a lack of significant sequence similarity. Recombinant subdomains of cf76 were challenged with survivor-cat antiserum and found to be highly seroreactive. Comparison of eleven geographically diverse samples from the south-central and southeastern USA demonstrated 91–100% amino acid sequence identity across cf76, including a high level of conservation in an immunogenic 226 amino acid (24 kDa) carboxyl terminal domain. Using in situ hybridization, transcription of cf76 was documented in the schizogenous stage of parasite replication, the life stage that is believed to be the most important for development of a protective immune response. Collectively, these data point to identification of the first potential vaccine candidate antigen for cytauxzoonosis. Further, our bioinformatic approach emphasizes the use of comparative genomics as an accelerated path to developing vaccines against experimentally intractable pathogens.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Novel Candidate Vaccine for Cytauxzoonosis Inferred from Comparative Apicomplexan Genomics

et al. 2013

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“…(Jefferies et al, 2007), Cytauxzoon felis (Birkenheuer et al, 2006) and Hepatozoon spp. (Ujvari et al, 2004) were used for DNA amplification (Table 1).…”

Section: Methodsmentioning

confidence: 99%

Tick-borne agents in domesticated and stray cats from the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil

André

Herrera

Fernandes

et al. 2015

Ticks and Tick-borne Diseases

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“…Polymerase Chain Reaction (PCR) was performed with primer pair specific for C. felis 18S rRNA gene, as previously described by Birkenheuer et al (2006). Both positive (known positive sample gently provided by Dr. Adivaldo Henrique da Fonseca, UFRRJ, RJ, Brazil) and negative (Ultra-Pure DNase/RNase-Free Distilled Water, Invitrogen, Carlsbad, CA, USA) controls were used in each set of reactions The 284 bp DNA fragment was purified using the PureLink PCR Purification Kit (Invitrogen, Carlsbad, CA, USA) and subjected to direct sequencing in both directions.…”

Section: Introductionmentioning

confidence: 99%

Cytauxzoon felis and 'Candidatus Mycoplasma haemominutum' coinfection in a Brazilian domestic cat (Felis catus)

Maia

Cerqueira

Macieira

et al. 2013

Rev. Bras. Parasitol. Vet.

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Cytauxzoon felis and 'Candidatus Mycoplasma haemominutum' coinfection in a Brazilian domestic cat (Felis catus) AbstractThis article describes the first detection of Cytauxzoon felis, using molecular techniques, in a naturally infected domestic cat from Brazil, South America. Coinfection with 'Candidatus Mycoplasma haemominutum' was also found. The molecular identification of the piroplasmid species was performed by Polymerase Chain Reaction (PCR) and sequencing analysis. A 284 pb fragment of the gene encoding the 18S ribosomal RNA region was amplified and showed 99% identity with other C. felis strains from North America. In addition, PCR-RFLP (restriction fragment length polymorphism) analysis, which amplifies a 595 bp fragment of the gene encoding 16S ribosomal RNA of some bacterial species, identified the co-infecting species as 'Candidatus M. haemominutum'.

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Development and evaluation of a PCR assay for the detection of Cytauxzoon felis DNA in feline blood samples

Cited by 47 publications

References 17 publications

A Novel Candidate Vaccine for Cytauxzoonosis Inferred from Comparative Apicomplexan Genomics

A Novel Candidate Vaccine for Cytauxzoonosis Inferred from Comparative Apicomplexan Genomics

Tick-borne agents in domesticated and stray cats from the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil

Cytauxzoon felis and 'Candidatus Mycoplasma haemominutum' coinfection in a Brazilian domestic cat (Felis catus)

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