Application of the polymerase chain reaction for the detection of Ehrlichia canis in tissues of dogs

Iqbal, Zafar; Rikihisa, Yasuko

doi:10.1016/0378-1135(94)90059-0

Cited by 31 publications

(34 citation statements)

References 14 publications

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“…Several studies of other Ehrlichia species have been performed by conventional onestep PCR. Iqbal and Rikihisa (14) reported that PCR can detect as little as 15 pg of DNA from purified Ehrlichia canis. Iqbal et al (13) reported that 20 pg of DNA extracted from E. canis-infected DH82 cells could be detected by PCR.…”

Section: Discussionmentioning

confidence: 99%

Sensitive Detection of Ehrlichia chaffeensis in Cell Culture, Blood, and Tick Specimens by Reverse Transcription-PCR

Felek¹,

Ünver²,

Stich

et al. 2001

J Clin Microbiol

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Ehrlichia chaffeensis is an obligatory intracellular bacterium of monocytes and macrophages and the etiologic agent of human monocytic ehrlichiosis, an emerging zoonosis. The Lone Star tick (Amblyomma americanum) has been implicated as the primary vector of E. chaffeensis. The present study examined the sensitivity of the nested reverse transcription (RT)-PCR based on the 16S rRNA gene relative to that of the nested PCR for detection of E. chaffeensis in infected DH82 cells, experimentally infected dog peripheral blood mononuclear cells, or experimentally infected A. americanum tick samples. The RT-PCR was found to be approximately 100 times more sensitive than the PCR for detection of E. chaffeensis regardless of the nature of the specimens. Thus, this RT-PCR is useful for detection of E. chaffeensis when a high sensitivity is required. Positive results by RT-PCR also imply the presence of viable pathogens. This is the first demonstration of RNA of E. chaffeensis in infected blood and acquisition-fed male, nymphal, and larval A. americanum ticks.

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Section: Discussionmentioning

confidence: 99%

Sensitive Detection of Ehrlichia chaffeensis in Cell Culture, Blood, and Tick Specimens by Reverse Transcription-PCR

Felek¹,

Ünver²,

Stich

et al. 2001

J Clin Microbiol

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“…This PCR/CH assay appears to be more sensitive than the previously reported PCR assay. 10 However, a direct comparison cannot be made because the sensitivity study in a previous report utilized DNA from infected cell culture cells, not genomic DNA from purified E. canis. In our investigation, PCR products were often not visualized in ethidium bromide-stained agarose gels but were detected with CH.…”

Section: Discussionmentioning

confidence: 99%

“…Alternatively, the use of cell culture isolation (CCI) to detect E. canis in blood from infected dogs is both sensitive and specific . 9,10 However, CCI requires 1-4 weeks to obtain results, thus limiting its usefulness as a rapid diagnostic tool.…”

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confidence: 99%

PCR Detection of Acute Ehrlichia Canis Infection in Dogs

et al. 1996

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Abstract.A polymerase chain reaction (PCR)-based detection assay that specifically detected Ehrlichia canis in dogs with acute infections was developed. A region of the 16S ribosomal RNA gene of E. canis was targeted for PCR amplification and chemiluminescent hybridization (CH) with a complementary internal 287-base pair (bp) oligonucleotide probe. The CH improved the PCR assay sensitivity 1,000-fold as compared with visualization on ethidium bromide-stained agarose gels. The PCR assay with CH (PCRKH) detected as little as 30 fg of E. canis genomic DNA, the equivalent of approximately 150 E. canis organisms. The 495-bp product defined by the specific primers was not detected when genomic DNA from E. platys, E. chaffeensis, E. risticii, and E. equi were used in the PCR/CH assay. The PCR/CH assay was tested with unfractionated blood samples collected from 9 dogs experimentally infected with E. canis. The PCR/CH assay had greater detection sensitivity than did cell culture isolation (CCI) from infected blood. PCR/CH detected E. canis 7 days prior to CCI in 4 of 6 experimentally infected dogs. The results obtained with the PCR/CH assay otherwise consistently matched the results obtained by CCI. This PCR/CH assay is a rapid, sensitive, and specific method for E. canis detection with sensitivity comparable to or exceeding that of CCI. A diagnosis of E. canis using this PCR/CH assay can be made in 2 days as compared with 14 weeks for CCI. The PCR/CH assay appears to be an acceptable alternative or complement to current diagnostic techniques.Canine ehrlichiosis is a tick-borne disease caused by an obligate intracellular parasite, Ehrlichia canis, which resides and replicates within mononuclear cells.7 The organism has a global distribution, including the United States, northern and southern African countries, Europe, Southeast Asia, and India.1,2,5-7 The association of E. canis with hemorrhagic disease in dogs was first described in 1969.8 Diagnosis of canine in the early stage of infection is important to ensure early treatment and a good prognosis.7 Dogs in the acute phase of the disease demonstrate dramatic improvement in hematologic and clinical responses 24-48 hours after therapy.7 However, dogs in the chronic stage of the disease have a poor prognosis and may not improve following treatment. 7,11Currently, definitive diagnosis of canine ehrlichiosis is based on hematologic, biochemical, and serologic findings.7 However, the most clinically reliable method for diagnosis of canine ehrlichiosis is by serologic techniques such as immunofluorescent antibody test (IFA) and plate latex agglutination that detect E. canis serum antibodies.7 However, serologic tests have several disadvantages: IgM and IgG antibodies are not detectable until at least 1-3 weeks postinfection, and cross-reactions occur between Ehrlichia species. Both of these factors can sometimes make a differential diagnosis difficult. Alternatively, the use of cell culture isolation (CCI) to detect E. canis in blood from infected dogs is both sensitive and ...

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“…It is a highly sensitive method for the early detection (usually 4-10 days post-inoculation), molecular characterization and quantifi cation (real-time PCR) of the ehrlichial organisms [1,62,75,76]. Also, PCR is more useful than serology, for the documentation of concurrent infections with different ehrlichial species and the post-treatment monitoring [5,6,77,78]. Importantly, in dogs with profound aplastic pancytopenia the diagnostic sensitivity of PCR may be suboptimal [16].…”

Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Canine Monocytic Ehrlichiosis: An Update on Diagnosis and Treatment

Mylonakis

Θεοδώρου

2017

Acta Veterinaria

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Canine monocytic ehrlichiosis (CME) is a tick-borne disease of worldwide distribution. The major causative agent is Ehrlichia canis, a gram-negative, obligate intracellular, pleomorphic bacterium of the genus Ehrlichia, which infects monocytes, macrophages and lymphocytes, forming intracytoplasmic, membrane-bound bacterial aggregates, called morulae. After an incubation period of 8-20 days, the course of E. canis infection, can be sequentially divided into acute, subclinical and chronic phases, although these phases can hardly be distinguished in the clinical setting. Clinical recovery is the typical outcome of acutely infected dogs, entering the subclinical phase, during which they show no or minimal clinical signs and/or mild hematological abnormalities. Immunocompetent dogs may eliminate the infection during the acute or subclinical phases, but an unpredictable proportion of dogs will eventually develop the chronic phase, characterized by aplastic pancytopenia and high mortality, due to septicemia and/or severe bleeding. This article outlines briefl y the pathogenesis of CME due to E. canis, and more thoroughly reviews the recent scientifi c literature pertaining to the diagnosis and treatment of this devastating disease.

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Application of the polymerase chain reaction for the detection of Ehrlichia canis in tissues of dogs

Cited by 31 publications

References 14 publications

Sensitive Detection of Ehrlichia chaffeensis in Cell Culture, Blood, and Tick Specimens by Reverse Transcription-PCR

Sensitive Detection of Ehrlichia chaffeensis in Cell Culture, Blood, and Tick Specimens by Reverse Transcription-PCR

PCR Detection of Acute Ehrlichia Canis Infection in Dogs

Canine Monocytic Ehrlichiosis: An Update on Diagnosis and Treatment

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