scholarly journals Demonstrating the presence of Ehrlichia canis DNA from different tissues of dogs with negative PCR in blood

2020 ◽  
Author(s):  
Carlos Arturo Rodríguez-Alarcón ◽  
Diana M. Beristain-Ruiz ◽  
Angélica Olivares-Muñoz ◽  
Andrés Quezada-Casasola ◽  
Federico Pérez-Casio ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cross Reactivity ◽  
Ehrlichia Canis ◽  
Morbidity And Mortality ◽  
Blood Samples ◽  
Serologic Tests ◽  
Negative Results ◽  
Target Organs ◽  
Positive Results ◽  
Different Tissues

Abstract Background: Nowadays, Ehrlichia canis receives more attention because of its great morbidity and mortality in animals. Dogs in the subclinical and chronic phases can be asymptomatic, and serologic tests show cross-reactivity and fail to differentiate between current and past infections. Moreover, there could be low parasitaemia, and E. canis might be found only in target organs, hence negative by PCR in blood. Methods: We evaluated by PCR the prevalence of E. canis in blood, liver, spleen, lymphatic nodules, and bone marrow in 59 recently euthanized dogs that had ticks but were clinically healthy. Results: In total, 52.55% of the blood PCRs for E. canis were negative, yet 61.30% yield positive results in tissue biopsies as follows: 63.15% from bone marrow, 52.63% from liver, 47.36% from spleen and 15.78% from lymphatic nodules. In addition, 33% had infection in three tissues (spleen, liver and bone marrow). Conclusions: Our results show prevalence of E. canis in tissue from dogs that were negative by PCR in blood. E. canis DNA in tissue was 30% lower in dogs that tested negative in blood samples by PCR, compared to those that were positive. However, it must be taken into account that some dogs with negative results were positive for E. canis in others tissues.

scholarly journals Demonstrating the presence of Ehrlichia canis DNA from different tissues of dogs with suspected subclinical ehrlichiosis

2020 ◽  
Author(s):  
Carlos Arturo Rodríguez-Alarcón ◽  
Diana M. Beristain-Ruiz ◽  
Angélica Olivares-Muñoz ◽  
Andrés Quezada-Casasola ◽  
Federico Pérez-Casio ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cross Reactivity ◽  
Ehrlichia Canis ◽  
Morbidity And Mortality ◽  
Blood Samples ◽  
Serologic Tests ◽  
Negative Results ◽  
Target Organs ◽  
Positive Results ◽  
Different Tissues

Abstract Background: Nowadays, Ehrlichia canis receives more attention because of its great morbidity and mortality in animals. Dogs in the subclinical and chronic phases can be asymptomatic, and serologic tests show cross-reactivity and fail to differentiate between current and past infections. Moreover, there could be low parasitaemia, and E. canis might be found only in target organs, hence negative by PCR in blood. Methods: We evaluated by PCR the prevalence of E. canis in blood, liver, spleen, lymphatic nodules, and bone marrow in 59 recently euthanized dogs that had ticks but were clinically healthy. Results: In total, 52.55% of the blood PCRs for E. canis were negative, yet 61.30% yield positive results in tissue biopsies as follows: 63.15% from bone marrow, 52.63% from liver, 47.36% from spleen and 15.78% from lymphatic nodules. In addition, 33% had infection in three tissues (spleen, liver and bone marrow). Conclusions: Our results show prevalence of E. canis in tissue from dogs that were negative by PCR in blood. E. canis DNA in tissue was 30% lower in dogs that tested negative in blood samples by PCR, compared to those that were positive. However, it must be taken into account that some dogs with negative results were positive for E. canis in others tissues.

scholarly journals Demonstrating the presence of Ehrlichia canis DNA from different tissues of dogs with suspected subclinical ehrlichiosis

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Carlos A. Rodríguez-Alarcón ◽  
Diana M. Beristain-Ruiz ◽  
Angélica Olivares-Muñoz ◽  
Andrés Quezada-Casasola ◽  
Federico Pérez-Casio ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Lymph Node ◽  
Cross Reactivity ◽  
Ehrlichia Canis ◽  
Serological Tests ◽  
Blood Samples ◽  
Negative Results ◽  
Target Organs ◽  
Polymerase Chain ◽  
Positive Results

Abstract Background Nowadays, Ehrlichia canis receives increasing attention because of its great morbidity and mortality in animals. Dogs in the subclinical and chronic phases can be asymptomatic, and serological tests show cross-reactivity and fail to differentiate between current and past infections. Moreover, there could be low parasitaemia, and E. canis might be found only in target organs, hence causing results to be negative by polymerase chain reaction (PCR) on blood samples. Methods We evaluated by PCR the prevalence of E. canis in blood, liver, spleen, lymph node and bone marrow samples of 59 recently euthanised dogs that had ticks but were clinically healthy. Results In total, 52.55% of the blood PCRs for E. canis were negative, yet 61.30% yielded positive results from tissue biopsies and were as follows: 63.15% from bone marrow; 52.63% from liver; 47.36% from spleen; and 15.78% from lymph node. In addition, 33% had infection in three tissues (spleen, liver and bone marrow). Conclusions Our results show the prevalence of E. canis from tissues of dogs that were negative by blood PCR. Ehrlichia canis DNA in tissue was 30% lower in dogs that tested negative in PCR of blood samples compared to those that were positive. However, it must be taken into account that some dogs with negative results were positive for E. canis in other tissues.

scholarly journals Demonstrating the presence of Ehrlichia canis DNA from different tissues of dogs with negative PCR in blood

2020 ◽  
Author(s):  
Carlos Arturo Rodríguez-Alarcón ◽  
Diana M. Beristain-Ruiz ◽  
Angélica Olivares-Muñoz ◽  
Andrés Quezada-Casasola ◽  
Federico Pérez-Casio ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Lymph Node ◽  
Cross Reactivity ◽  
Ehrlichia Canis ◽  
Chain Reaction ◽  
Blood Samples ◽  
Negative Results ◽  
Target Organs ◽  
Polymerase Chain ◽  
Positive Results

Abstract Background: Nowadays, Ehrlichia canis receives increasing attention because of its great morbidity and mortality in animals. Dogs in the subclinical and chronic phases can be asymptomatic, and serologic tests show cross-reactivity and fail to differentiate between current and past infections. Moreover, there could be low parasitaemia, and E. canis might be found only in target organs, hence causing results to be negative by polymerase chain reaction (PCR) on blood samples.Methods: We evaluated by PCR the prevalence of E. canis in blood, liver, spleen, lymph node and bone marrow of 59 recently euthanised dogs that had ticks but were clinically healthy.Results: In total, 52.55% of the blood PCRs for E. canis were negative, yet 61.30% yielded positive results from tissue biopsies and were as follows: 63.15% from bone marrow, 52.63% from liver, 47.36% from spleen and 15.78% from lymph node. In addition, 33% had infection in three tissues (spleen, liver and bone marrow).Conclusions: Our results show the prevalence of E. canis from tissues of dogs that were negative by blood PCR. E. canis DNA in tissue was 30% lower in dogs that tested negative in PCR of blood samples compared to those that were positive. However, it must be taken into account that some dogs with negative results were positive for E. canis in other tissues.

scholarly journals Demonstrating the presence of Ehrlichia canis DNA from different tissues of dogs with negative PCR in blood

2020 ◽  
Author(s):  
Carlos Arturo Rodríguez-Alarcón ◽  
Diana M. Beristain-Ruiz ◽  
Angélica Olivares-Muñoz ◽  
Andrés Quezada-Casasola ◽  
Federico Pérez-Casio ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Lymph Node ◽  
Cross Reactivity ◽  
Ehrlichia Canis ◽  
Serological Tests ◽  
Blood Samples ◽  
Negative Results ◽  
Target Organs ◽  
Polymerase Chain ◽  
Positive Results

Abstract Background: Nowadays, Ehrlichia canis receives increasing attention because of its great morbidity and mortality in animals. Dogs in the subclinical and chronic phases can be asymptomatic, and serological tests show cross-reactivity and fail to differentiate between current and past infections. Moreover, there could be low parasitaemia, and E. canis might be found only in target organs, hence causing results to be negative by polymerase chain reaction (PCR) on blood samples.Methods: We evaluated by PCR the prevalence of E. canis in blood, liver, spleen, lymph node and bone marrow samples of 59 recently euthanised dogs that had ticks but were clinically healthy.Results: In total, 52.55% of the blood PCRs for E. canis were negative, yet 61.30% yielded positive results from tissue biopsies and were as follows: 63.15% from bone marrow; 52.63% from liver; 47.36% from spleen; and 15.78% from lymph node. In addition, 33% had infection in three tissues (spleen, liver and bone marrow).Conclusions: Our results show the prevalence of E. canis from tissues of dogs that were negative by blood PCR. Ehrlichia canis DNA in tissue was 30% lower in dogs that tested negative in PCR of blood samples compared to those that were positive. However, it must be taken into account that some dogs with negative results were positive for E. canis in other tissues.

scholarly journals Antibodies produced by dogs successfully challenged with live Leptospira spp. did not cross-react to Brucella antigen in a commercial rapid slide agglutination test that detects antibodies to Brucella canis

2018 ◽  
Vol 31 (1) ◽  
pp. 83-85
Author(s):  
Matthew R. Krecic
Keyword(s):  
Cross Reactivity ◽  
Agglutination Test ◽  
Slide Agglutination ◽  
Serologic Tests ◽  
Experimental Infections ◽  
Negative Results ◽  
Brucella Canis ◽  
Slide Agglutination Test ◽  
Leptospira Spp ◽  
Canine Brucellosis

Brucella canis is a cause of canine infertility and abortion. Veterinarians and veterinary laboratorians screen for antibodies to B. canis with serologic tests including a rapid slide agglutination test (RSAT; D-Tec CB, Zoetis, San Diego, CA). False-positive results are possible because of cross-reactivity to antibodies to some gram-negative bacteria. Cross-reactivity has been reported between antibodies of Brucella abortus and Leptospira spp. with serologic tests for bovine brucellosis; however, this has not been documented with serologic tests for canine brucellosis, to the author’s knowledge. The RSAT was evaluated with the sera from dogs experimentally challenged with 1 of 4 serovars of Leptospira spp.: L. kirschneri serovar Grippotyphosa, or L. interrogans serovars Canicola, Icterohaemorrhagiae, or Pomona. Experimental infections were confirmed through results of microscopic agglutination testing and/or lateral flow immunochromatography testing. The sera of 32 dogs collected at day 0 and days 7, 10, and 14 yielded negative results with the RSAT. Antibodies produced through experimental infections to these 4 serovars of Leptospira spp. did not cross-react with Brucella antigen with the RSAT; therefore, cross-reactivity of anti-leptospiral antibodies may not be of concern for B. canis rapid slide agglutination testing of dogs.

scholarly journals Short Communication: Molecular characterization and blood hematology profile of dogs infected by Ehrlichia canis in Yogyakarta, Indonesia

2020 ◽  
Vol 21 (7) ◽  
Author(s):  
Yustinus oswin primajuni Wuhan ◽  
Aris Haryanto ◽  
Ida Tjahajati
Keyword(s):  
Molecular Characterization ◽  
Microscopic Examination ◽  
Short Communication ◽  
Pcr Amplification ◽  
Ehrlichia Canis ◽  
Infected Blood ◽  
Blood Samples ◽  
Glta Gene ◽  
Vector Borne ◽  
Positive Results

Abstract. Wuhan YOP, Haryanto A, Tjahajati I. 2020. Short Communication: Molecular characterization and blood hematology profile of dogs infected by Ehrlichia canis in Yogyakarta, Indonesia. Biodiversitas 21: 3242-3248. Ehrlichia canis is Gram-negative intracellular obligate bacteria that cause ehrlichiosis, a companion vector-borne disease is a potentially fatal disease that attacks dogs. The purpose of this study was to molecular characterize and determine the features of Ehrlichia-infected blood based on the amplification of the gltA gene in Ehrlichia infected dogs from Yogyakarta, Indonesia. Blood samples were collected from 51 dog patients from the Prof. Dr. Soeparwi Animal Hospital, animal clinics, and pet shops based on the anamnesis, clinical sign, and physical examination, followed by microscopic examination, routine hematology, PCR amplification, and DNA sequencing were carried out on the blood samples. Based on positive PCR amplification and blood hematology profile examination ehrlichiosis-positive in dogs showed that thrombocytopenia case was 82.3%, anemia was 70.5%, eosinopenia was 70.5%, neutropenia was 29.4%, monocytopenia was 23%, leukopenia was 17% and lymphopenia was 11.7%. Morulae of Ehrlichia sp.was not found in microscopic examination. Molecularly, detected of E. canis using the gltA gene showed that 34% of samples were positive results. Then 5 of positive Ehrlichia samples were DNA sequenced, they showed a high homology of 100% with Hat Yai isolates (KU765199.1). There was no genetic diversity between E. canis samples in Yogyakarta.

scholarly journals VECTOR-BORNE PATHOGENS-TESTING IN A ROMANIAN CANINE BLOOD BANK

2018 ◽  
Vol 6 ◽  
pp. 904-911
Author(s):  
Teodor-Stefan Ionescu ◽  
Sinziana Radulescu ◽  
Ruxandra Florea ◽  
Stelian Baraitareanu ◽  
Doina Danes
Keyword(s):  
Blood Donors ◽  
Holistic Approach ◽  
Blood Bank ◽  
Focused Attention ◽  
Donor Pool ◽  
Blood Samples ◽  
Negative Results ◽  
Blood Banks ◽  
Vector Borne ◽  
Positive Results

INTRODUCTION: Canine blood banking in veterinary medicine is an expanding market. Once the demand for blood products increased all over the world, canine blood banks have focused attention on the risk of spreading diseases through blood transfused products. The need to preserve a healthy donor-pool, free of blood-borne infectious diseases, mainly in endemic areas, led to the implementation of appropriate protocols for screening canine blood donors using specific tests.OBJECTIVES: The aim of this study was to evaluate the presence of Anaplasma phagocytophilum/Anaplasmaplatys, Echrlichiacanis/Echrlichiaewingii, Dirofilariaimmitis and Borrelia burgdorferi using the enzyme immunoassay technology (EIA) among the donors of a Romanian canine blood bank, from January 2015 to December 2016.METHODS: Blood samples from 575 donors were collected and 1253 tests were performed with SNAP 4DX Plus® (IDEXX Laboratories, Fremont, CA) to reveal the presence of D. immitis antigens and the antibodies toward A. phagocytophilum and/or A. platys, E. canis and/or E. ewingii and B. burgdorferi.RESULTS: The results of this holistic approach show that all blood samples provided negative results for B. burgdorferi and E.canis/E. ewingii (0/1253), while 0.87% (11/1253) samples provided positive results for A. phagocytophilum/A. platys and 6.94% (87/1253) for D. immitis.CONCLUSION: The next studied topic would be to compare the results provided by the EIA technology with results of real time PCR and qPCR regarding these vector-borne pathogens.

scholarly journals Ehrlichia canis detection in dogs from Várzea Grande: a comparative analysis of blood and bone marrow samples

2015 ◽  
Vol 46 (2) ◽  
pp. 310-314 ◽  
Author(s):  
Herica Makino ◽  
Valéria Régia Franco Sousa ◽  
Mahyumi Fujimori ◽  
Juliana Yuki Rodrigues ◽  
Alvaro Felipe Lima Ruy Dias ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Clinical Signs ◽  
Cross Sectional Study ◽  
Buffy Coat ◽  
Ehrlichia Canis ◽  
Sectional Study ◽  
Cross Sectional ◽  
Mato Grosso ◽  
Blood Samples ◽  
Canine Ehrlichiosis

ABSTRACT: The objective of this study was to compare the DNA detection of Ehrlichia canis in blood and bone marrow to determine the prevalence of the agent in Várzea Grande, Mato Grosso. Blood samples and bone marrow from 80 dogs of both sexes, different breeds and age, were collected and processed for a cross-sectional study performed using nested PCR. Of the 80 dogs, 61 (76.3%) had E. canis DNA in one of the samples. The buffy coat was positive in 42 dogs (52.5%) and the bone marrow was positive in 33 (41.3%). There was no significant association between the positive biological samples of either the buffy coat or bone marrow and the presence or absence of clinical signs (P=0.49). No risk factor was associated with infection in the studied area. The bone marrow samples were efficient for the molecular diagnosis of canine ehrlichiosis, particularly when there was a negative blood sample, although infection was present.

Improving the Sensitivity and the Specificity of HLA-B27 Typing by Replacing Serological Tests with a TaqMan-PCR Assay

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4720-4720
Author(s):  
Xiaoduan Weng ◽  
Louise Robin ◽  
Marie-lise Audet ◽  
Linda Hébert ◽  
Ginette Lapointe ◽  
...  
Keyword(s):  
Detection System ◽  
Cross Reactivity ◽  
Hla B27 ◽  
Serological Tests ◽  
Dna Assay ◽  
Negative Results ◽  
Flow Cytometric ◽  
Antigen Assay ◽  
Taqman Pcr ◽  
Positive Results

Abstract HLA-B27 is a strong diagnostic biomarker as it is found in 90– 95% of ankylosing spondylitis. Routinely, the false-positive results generated by cross-reactivity of HLA-B27 monoclonal antibodies (mAbs) with some other type of HLA-B antigens are problematic. The present study aims at improving the sensitivity and specificity of typing for HLA-B27 by using a more accurate DNA assay. A real time sequence specific primer polymerase chain reaction (PCR-SSP) is performed by using MGB TaqMan oligoprobe and an ABI PRISM™ 7500 sequence detection system. The TaqMan PCR-SSP assay is compared with Immunophenotyping by flow cytometric antigen assay (BD HLA-B27-Kit, clone GS145.2). The results are finally confirmed by sequencing (ABI PRISM ® 3100). As summarised in the table, three methods are identical for clearly positive and negative results. There is 90% concordance for borderline negative results obtained by immunophenotyping with TaqMan PCR-SSC and/or sequencing. However, only 15.4% of borderline positives obtained by immunophenotyping are confirmed as true HLA-B27 positives by sequencing as well as TaqMan PCR-SSC. TaqMan PCR-SSP DNA assay completely correlates with sequencing (gold standard) with 100% PPV and NPV, compared to a PPV of 15% and a NPV of 98% for borderline results by immunophenotyping. Conclusion: when compared with flow cytometric antigen assay, typing HLA-B27 by TaqMan PCR-SSP clearly demonstrates an advantage to better establish the genotype of the patient. Specifically, there is a marked difference for ambitious positive results from immunophenotyping. Moreover, compared to immunophenotyping, there is no need of fresh samples, can test a larger number of samples concomitantly, and reduces technical time as well as cost. N = 52 Immunophenotyping TaqMan PCR-SSP Sequencing Immuno vs Sequencing TaqMan PCR-SSP vs Sequencing Positive 11 11 11 100% 100% Negative 17 17 17 100% 100% Positive (borderline by Immunophenotyping) 13 2 2 15% 100% Negative (borderline ) by Immunophenotyping) 10 9 9 98% 100%

Critical Evaluation of Impedance Phlebography for the Diagnosis of Deep Vein Thrombosis

1974 ◽  
Vol 31 (02) ◽  
pp. 273-278
Author(s):  
Kenneth K Wu ◽  
John C Hoak ◽  
Robert W Barnes ◽  
Stuart L Frankel
Keyword(s):  
Deep Vein Thrombosis ◽  
False Positive ◽  
False Negative ◽  
Critical Evaluation ◽  
Original Method ◽  
Vein Thrombosis ◽  
Negative Results ◽  
False Negative Results ◽  
Deep Vein ◽  
Positive Results

SummaryIn order to evaluate its daily variability and reliability, impedance phlebography was performed daily or on alternate days on 61 patients with deep vein thrombosis, of whom 47 also had 125I-fibrinogen uptake tests and 22 had radiographic venography. The results showed that impedance phlebography was highly variable and poorly reliable. False positive results were noted in 8 limbs (18%) and false negative results in 3 limbs (7%). Despite its being simple, rapid and noninvasive, its clinical usefulness is doubtful when performed according to the original method.

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