Use of an automated system for detection of canine serum antibodies against <i>Ehrlichia canis</i> glycoprotein 36

Moroff, Scott; Sokolchik, Irene; Woodring, Todd; Woodruff, Colby; Atkinson, Brett; Lappin, Michael R.

doi:10.1177/1040638714534849

Cited by 7 publications

(7 citation statements)

References 17 publications

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“…This may be due to pancytopenia and bone marrow failure (Saito and Walker, 2016). E. canis DNA in infected dog blood can be detected earlier than serum antibodies, while the latter continue to persist even after doxycycline therapy (Moroff et al, 2014). Moreover, cross reactivity between different Ehrlichia spp.…”

Section: Discussionmentioning

confidence: 99%

Canine Monocytic Ehrlichiosis among working dogs of organised kennels in India: A comprehensive analyses of clinico-pathology, serological and molecular epidemiological approach

Mittal

Kundu

Chakravarti

et al. 2017

Preventive Veterinary Medicine

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Canine Monocytic Ehrlichiosis (CME) is a serious tick-borne rickettsial disease affecting canine populations globally. Besides few reports from stray and pet dogs from localised geographical regions (cities/towns/small states), a comprehensive study on prevalence of Ehrlichia canis (E. canis) among working dogs from different geo-climatic zones of India was pertinently lacking. Study of CME among these dog populations was thus carried out, encompassing clinical aspects and different diagnostic methodologies viz., microscopy, serology and molecular biology. During the two-year study period, clinical specimens from 225 cases suspected of canine ehrlichiosis were examined for clinical pathology and presence of the haemoparasites. Overall prevalence of ehrlichiosis by microscopic examination, commercial dot-ELISA kit and nested PCR assay was estimated to be 1.3%, 19.1% and 5.8%, respectively, which were found to be statistically significant by McNemar Chi squared test (p<0.05). It was also observed that possibly due to widespread use of doxycycline therapy in field, CME presently does not remain a potential threat which it uses to pose earlier. However, concurrent infections of E. canis and Babesia gibsoni were found to be mostly fatal. Keeping in view of high number of apparently healthy dogs (24) out of total positive cases (46) observed during the study, it is recommended that prevalence studies on CME should also involve screening of apparently healthy dogs. Phylogenetic analysis carried on partial sequencing of 16S rRNA of E. canis strains revealed that all of the Indian strains clustered in a single clade with other E. canis species from India and rest of the world. Molecular divergence was observed among the sequences of Brazilian and American isolates which were also included in the present study. These findings have thus opened a new paradigm for planning of pragmatic control strategies against CME.

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Section: Discussionmentioning

confidence: 99%

Canine Monocytic Ehrlichiosis among working dogs of organised kennels in India: A comprehensive analyses of clinico-pathology, serological and molecular epidemiological approach

Mittal

Kundu

Chakravarti

et al. 2017

Preventive Veterinary Medicine

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“…Currently, serological diagnostic tests for canine monocytic ehrlichiosis include IFAT, ELISA, and in-house commercial test kits commonly used by routine veterinary medicine practices. Specific IgG against E. canis in the suspected dog's sera or whole blood is a primary target of these serological tests ( Cárdenas et al., 2007 ; Baneth et al., 2009 ; Moroff et al., 2014 ). The experimental infections by Baneth et al.…”

Section: Introductionmentioning

confidence: 99%

Detection of specific IgM and IgG antibodies in acute canine monocytic ehrlichiosis that recognize recombinant gp36 antigens

Kaewmongkol

Suwan

Sirinarumitr

et al. 2020

Heliyon

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The efficacy of antibody detection tools for all stages of Ehrlichia canis infections and for various genotypes remains unclear. We produced recombinant gp36 (rgp36) antigens from different isolates of Thai E. canis to confirm the immunoreactivities to these recombinant proteins from naturally infected dogs. Sera and blood samples were taken from 21 dogs naturally infected with E. canis and in the clinical stages of acute phase ehrlichiosis. The expression vectors and competent E. coli produced two isolates of rgp36. These two major rgp36s were recognized by the dogs' sera in Western blotting, with both anti-dog IgM and IgG used as secondary antibodies. The two different genotypes of these local recombinant immunoreactive proteins were gp36 subgroup A (isolate 1055) and subgroup B (isolate 533). The Western blot analyses successfully identified both specific IgM and IgG from the dogs' sera. Of all 21 cases, five dogs presented specific IgM, twenty dogs presented specific IgG, and the commercial test used found fifteen seropositive dogs. There were four dogs that presented both specific IgM and IgG. Only one dog presented specific IgM only. This report is the first identification of a specific IgM in dogs in response to acute infections with E. canis . The recombinant gp36 isolates may be useful as potential antigenic material for subsequent serological tests that have a high possibility for differentiating between acute, chronic, primary, and nonprimary infections with E. canis .

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“…15 Positive and negative control sera used in assay titrations were obtained from the dogs in experiment 2 before (negative control) and after (positive control) infestation with I. scapularis. The optimal peptide concentrations were determined by assessing optimal signal-to-noise ratio, varying printed target concentrations, and buffer compositions.…”

Section: The Accuplex4 Systemmentioning

confidence: 99%

“…The use of the assay for detection of antibodies against peptides of Anaplasma phagocytophilum and Ehrlichia canis was previously reported. 14,15 The primary objective of the current study was to optimize detection of serum antibodies against multiple natural and artificial peptides of B. burgdorferi for use on the Accuplex4 system platform. The discs also contain peptides for detection of antibodies against natural and artificial peptides of A. phagocytophilum and E. canis, as well as an antigen of Dirofilaria immitis.…”

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confidence: 99%

“…17 The commercial automated fluorescent system, the Accuplex4 BioCD system, c used in our study can be used to assess antibody responses to multiple antigens using 70 µL of serum. 14,15 The system is a novel, silicon disc substrate multiplex platform for detection of peptides and antibodies, using a single serum or plasma sample. The use of the assay for detection of antibodies against peptides of Anaplasma phagocytophilum and Ehrlichia canis was previously reported.…”

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confidence: 99%

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Multiple antigen target approach using the Accuplex4 BioCD system to detect Borrelia burgdorferi antibodies in experimentally infected and vaccinated dogs

et al. 2015

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Abstract. The primary objective of our study was to optimize detection of serum antibodies to Borrelia burgdorferi using a new commercial automated fluorescence system (Accuplex4 BioCD system, Antech Diagnostics, Lake Success, New York). The system used multiple natural and artificial peptides-outer surface proteins (OspA, OspC, OspF), an outer membrane protein (P39), and a proprietary synthetic peptide (small Lyme peptide [SLP])-and the results were compared with a commercially available enzyme-linked immunosorbent assay that uses a proprietary peptide (C6). Sera from 4 groups were evaluated: dogs vaccinated with 1 of 3 commercially available vaccines (n = 18); dogs infested with adult Ixodes scapularis (black-legged tick; n = 18); dogs previously vaccinated and then infested with I. scapularis (n = 18); and dogs with B. burgdorferi infection that were then vaccinated (n = 14). All of the vaccines evaluated induced OspA responses. However, antibodies against OspF or C6 were not induced in any of the vaccinated dogs. Additionally, the OspF antibodies had 100% sensitivity and specificity when compared to antibodies against C6 peptide. In B. burgdorferi-infected dogs, antibodies against OspC and SLP were detected in serum sooner than antibodies against the other targets. Low levels of antibodies against OspA developed in 6 of 14 B. burgdorferi-infected, unvaccinated dogs and had the shortest duration compared to the other antibodies. Detection of antibody responses to multiple B. burgdorferi targets with this system can be used to help differentiate vaccinated dogs from exposed dogs as well as acute infection from chronic infection.

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Use of an automated system for detection of canine serum antibodies against Ehrlichia canis glycoprotein 36

Cited by 7 publications

References 17 publications

Canine Monocytic Ehrlichiosis among working dogs of organised kennels in India: A comprehensive analyses of clinico-pathology, serological and molecular epidemiological approach

Canine Monocytic Ehrlichiosis among working dogs of organised kennels in India: A comprehensive analyses of clinico-pathology, serological and molecular epidemiological approach

Detection of specific IgM and IgG antibodies in acute canine monocytic ehrlichiosis that recognize recombinant gp36 antigens

Multiple antigen target approach using the Accuplex4 BioCD system to detect Borrelia burgdorferi antibodies in experimentally infected and vaccinated dogs

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