Molecular Detection of<i>Anaplasma Platys</i>in Dogs Using Polymerase Chain Reaction and Reverse Line Blot Hybridization

Sparagano, Olivier; Vos, A.P. de; Paoletti, Barbara; Cammà, Cesare; Santis, Paola De; Otranto, Domenico; Giangaspero, Annunziata

doi:10.1177/104063870301500604

Cited by 46 publications

(32 citation statements)

References 27 publications

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“…The E. canis seroprevalence (i.e., 17.6%) recorded at the third follow-up time period was similar to the seroprevalence reported in another study of kenneled dogs from southern Italy (i.e., 14.9%) (31). However, the PCR prevalence of A. platys infection (from 30.4 to 57.7%) in naive dogs was higher than the prevalence previously reported in kenneled dogs from central (23%) (38) or southern (11.3%) (31) Italy. The higher prevalence in this study was potentially due to the incorporation of three different diagnostic testing results for each dog at each time point.…”

Section: Discussionsupporting

confidence: 85%

Diagnosis of Canine Vector-Borne Diseases in Young Dogs: a Longitudinal Study

Otranto¹,

Testini²,

Dantas‐Torres³

et al. 2010

J Clin Microbiol

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Canine vector-borne diseases (CVBDs) pose a diagnostic challenge, particularly when a dog is coinfected with more than one pathogen. The purpose of this study was to generate information about the diagnosis of CVBDs in young dogs following their first exposure to flea, tick, sand fly, louse, and mosquito vectors. From March 2008 to May 2009, 10 purpose-bred young naive beagle dogs and a cohort of 48 mixed-breed dogs living in an area to which CVBD is endemic in southern Italy were monitored using different diagnostic tests (cytology, serology, and PCR). Overall, PCR detected the highest number of dogs infected with Anaplasma platys, Babesia vogeli, and Ehrlichia canis, whereas seroconversion was a more sensitive indicator of exposure to Leishmania infantum. For A. platys infection, combining blood and buffy coat cytology in parallel enhanced the relative sensitivity (SE rel ) (87.3%). For B. vogeli, the best diagnostic combination was buffy coat cytology and serology used in parallel (SE rel , 67.5%), whereas serology and PCR used in parallel (SE rel , 100%) was the best combination for L. infantum. Overall, 12 (20.7%) dogs were coinfected; however, the percentage of new coinfections decreased from baseline (50%) to the first (33.3%) and second (16.6%) follow-up time points. Numbers of coinfections with A. platys and B. vogeli were significantly higher (P < 0.05) than coinfections with other pathogen combinations. The data generated in this study provide insights on the incidence of certain pathogens infecting young dogs in southern Italy, highlight important diagnostic testing limitations, and support the use of multiple diagnostic modalities when attempting to confirm a tick-borne infection in an individual dog or in a canine population.

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Section: Discussionsupporting

confidence: 85%

Diagnosis of Canine Vector-Borne Diseases in Young Dogs: a Longitudinal Study

Otranto¹,

Testini²,

Dantas‐Torres³

et al. 2010

J Clin Microbiol

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“…Collectively, these findings could support the presence of a unique or new Ehrlichia species on the island of Mallorca. Anaplasma platys infection is common in dogs living in the Mediterranean basin in countries such as Spain [33], Greece and Italy [39]. However, since A. platys does not serologically cross-react with Ec [11], it is unlikely that our Ec IFA results are due to A. platys infection.…”

Section: Discussionmentioning

confidence: 99%

A serological study of exposure to arthropod-borne pathogens in dogs from northeastern Spain

Solano‐Gallego

Llull

Osso³

et al. 2006

Vet. Res.

105

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-There is limited information regarding the prevalence of many vector borne pathogens in Europe and especially in Spanish dogs. We investigated 206 sick and 260 clinically healthy dogs from three different regions in northeastern Spain for antibodies to Rickettsia conorii (Rc), Ehrlichia canis (Ec), Anaplasma phagocytophilum (Ap), Bartonella henselae (Bh), Bartonella vinsonii subsp. berkhoffii (Bvb), Leishmania infantum (Li) and Borrelia burgdorferi (Bb) and for antigen of Dirofilaria immitis (Di). Total prevalences were the following: Rc (56.4%), Li (30%), Ec (16.7%), Bh (16.8%), Ap (11.5%), Bvb (1.07%), Di (0.6%) and Bb (0.6%). Seroprevalences for Rc, Ec, Ap, Bh, and Bvb and Bb and Di antigens were similar among the three different study sites. The Ec seroprevalence, as determined by Snap 3DX, was statistically lower in dogs from Mallorca (0%) than Tarragona (16%) and Barcelona (5%) (P < 0.0001). Detection of Rc antibodies was associated with seroreactivity to Ec and Ap antigens (P = 0.018 and P = 0.002, respectively). IFA Ec antibodies were associated with Ap seroreactivity (P < 0.0001). There was no association between the clinical status, sex, time of the year when samples were collected, life-style or exposure to fleas or ticks and a positive test result for Ec, Bh, Bvb, or Bb antibodies or Di antigens. Li seroreactivity was associated with illness and living outdoors (P < 0.0001, P = 0.029; respectively), Rc seroreactivity with the male gender (P = 0.028) and Ap seroreactivity with living outdoors (P = 0.045). This study indicates that exposure to Rc, Li, Ec or related Ehrlichia spp., Bh and Ap or a related spp., is common whereas Di, Bb and Bvb is uncommon among dogs from the Mediterranean basin. We also provide serological data that suggests the existence of a novel Ehrlichia species on Mallorca island. rickettsial pathogens / dogs / Spain

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“…30,[32][33][34][35][36][37][38][39] A PCR followed by the reverse line blot hybridization assay to detect species-specific amplified products is described recently for the simultaneous detection of Ehrlichia and Anaplasma species in ruminants and dogs. 40,41 In this assay, a 16S rRNA gene segment was amplified with a genera-specific primer set, and the products are hybridized to a membrane coated with species-specific oligonucleotide probes. The PCR assays described in the literature require additional protocols after a PCR or RT-PCR step.…”

Section: Discussionmentioning

confidence: 99%

Multiplex Detection of Ehrlichia and Anaplasma Species Pathogens in Peripheral Blood by Real-Time Reverse Transcriptase-Polymerase Chain Reaction

Sirigireddy

Ganta

2005

The Journal of Molecular Diagnostics

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Tick-borne infections are responsible for many emerging diseases in humans and several vertebrates. These include human infections with Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Ehrlichia ewingii. Because single or co-infections can result from tick bites, the availability of a rapid, multiplex molecular test will be valuable for timely diagnosis and treatment. Here, we describe a multiplex molecular test that can detect single or co-infections with up to five Ehrlichia and Anaplasma species. The test protocol includes the magnetic capture-based purification of 16S ribosomal RNA, its enrichment, and specific-pathogen(s) detection by real-time reverse transcriptase-polymerase chain reaction. We also report a unique cloning strategy to develop positive controls in the absence of a pathogen's genomic DNA. The test was assessed by examining blood samples from dogs suspected to be positive for ehrlichiosis. The dog was chosen as the model system because it is susceptible to acquire infections with up to five pathogens of the genera Ehrlichia and Anaplasma. The test identified single infections in the canine host with E. chaffeensis, E. canis, E. ewingii, A. phagocytophilum, and A. platys and co-infection with E. canis and A. platys. The multipathogen detection and novel positive control development procedures described here will be valuable in monitoring infections in people, other vertebrates, and ticks. (J Mol Diagn 2005, 7:308 -316) Several rickettsial agents of the family Anaplasmataceae cause severe, tick-borne pathogen infections in a wide range of vertebrate host species.

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Molecular Detection ofAnaplasma Platysin Dogs Using Polymerase Chain Reaction and Reverse Line Blot Hybridization

Cited by 46 publications

References 27 publications

Diagnosis of Canine Vector-Borne Diseases in Young Dogs: a Longitudinal Study

Diagnosis of Canine Vector-Borne Diseases in Young Dogs: a Longitudinal Study

A serological study of exposure to arthropod-borne pathogens in dogs from northeastern Spain

Multiplex Detection of Ehrlichia and Anaplasma Species Pathogens in Peripheral Blood by Real-Time Reverse Transcriptase-Polymerase Chain Reaction

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