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 ◽  
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 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 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.

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%

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.

Detection of Subclinical Systemic Disease in Primary CNS Lymphoma by Polymerase Chain Reaction of the Rearranged Immunoglobulin Heavy-Chain Genes

2006 ◽  
Vol 24 (29) ◽  
pp. 4754-4757 ◽  
Author(s):  
Kristoph Jahnke ◽  
Michael Hummel ◽  
Agnieszka Korfel ◽  
Thomas Burmeister ◽  
Philipp Kiewe ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Systemic Disease ◽  
Primary Cns Lymphoma ◽  
Cns Lymphoma ◽  
Chain Reaction ◽  
Blood Samples ◽  
Pcr Products ◽  
Polymerase Chain

Purpose To search for subclinical systemic disease in bone marrow and peripheral blood in patients with primary CNS lymphoma (PCNSL) to elucidate whether extracerebral relapse may represent a sequel of initial occult systemic disease rather than true extracerebral spread. Patients and Methods Bone marrow and peripheral-blood specimens of 24 PCNSL patients were examined using polymerase chain reaction (PCR) for analysis of clonally rearranged immunoglobulin heavy-chain (IgH) genes. Results Identical dominant PCR products were found in bone marrow aspirates, blood samples, and tumor biopsy specimens of two patients, indicating that the same tumor cell population is present in the CNS and in extracerebral sites. Follow-up IgH PCR performed in one of these patients in complete remission 24 months after diagnosis yielded a persistent monoclonal product in the blood. An oligoclonal IgH rearrangement pattern was found in the tumor specimen of two other patients, whereas bone marrow and blood samples demonstrated the same dominant PCR products. Follow-up PCR showed a persistent monoclonal amplificate in blood in one of these patients 27 months after diagnosis. Conclusion It could be demonstrated for the first time that subclinical systemic disease can be present in PCNSL patients at initial diagnosis. Our findings may have an impact on the understanding of PCNSL pathogenesis and the extent of staging and treatment.

scholarly journals Detection and molecular characterisation of Ehrlichia canis in naturally infected dogs in South West Nigeria

2018 ◽  
Vol 66 (1) ◽  
pp. 85-95 ◽  
Author(s):  
Olukayode Olugbenga Daramola ◽  
Michael Irewole Takeet ◽  
Ibironke Kofoworola Oyewusi ◽  
Mufutau Atanda Oyekunle ◽  
Adewale Oladele Talabi
Keyword(s):  
Ehrlichia Canis ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Rickettsial Disease ◽  
Blood Samples ◽  
Canine Ehrlichiosis ◽  
South West ◽  
Polymerase Chain ◽  
Reference Sequences ◽  
The 16S Rrna Gene

Canine ehrlichiosis is an important tick-borne rickettsial disease mainly caused by Ehrlichia canis. This study aimed to detect and characterise E. canis in dogs in Abeokuta, Nigeria by microscopy and nested PCR. Blood samples were collected from 205 dogs, thin smears were made, field-stained, and DNA was extracted from the blood samples. A partial region of the 16S rRNA gene was amplified by polymerase chain reaction (PCR) and sequenced unidirectionally. Ehrlichial morulae were detected in three dogs (1.5%). The PCR test revealed that 47 dogs (22.9%) were positive for E. canis. The lengths of the sequences obtained range from 374 bp to 376 bp with an average G-C content of 37% and 98–99% homology with the reference sequences in GenBank. The aligned autochthonous sequences were less polymorphic. The phylogenetic analysis separated sequences reported previously in Nigeria from the autochthonous sequences. The present work shows that the strain of E. canis detected in the study area is genetically different from those reported in the northern part of Nigeria and more closely related to sequences from Brazil and India.

scholarly journals False Positive Results in SARS-CoV-2 Serological Tests for Samples From Patients With Chronic Inflammatory Diseases

2021 ◽  
Vol 12 ◽  
Author(s):  
Nastya Kharlamova ◽  
Nicky Dunn ◽  
Sahl K. Bedri ◽  
Svante Jerling ◽  
Malin Almgren ◽  
...  
Keyword(s):  
Lupus Erythematosus ◽  
False Positive ◽  
Inflammatory Diseases ◽  
Serological Tests ◽  
Chronic Inflammatory Diseases ◽  
Negative Results ◽  
False Positive Signal ◽  
Lateral Flow Assays ◽  
Positive Results ◽  
Multiplex Bead

Patients with chronic inflammatory diseases are often treated with immunosuppressants and therefore are of particular concern during the SARS-CoV-2 pandemic. Serological tests will improve our understanding of the infection and immunity in this population, unless they tests give false positive results. The aim of this study was to evaluate the specificity of SARS-Cov-2 serological assays using samples from patients with chronic inflammatory diseases collected prior to April 2019, thus defined as negative. Samples from patients with multiple sclerosis (MS, n=10), rheumatoid arthritis (RA, n=47) with or without rheumatoid factor (RF) and/or anti-cyclic citrullinated peptide antibodies (anti-CCP2) and systemic lupus erythematosus (SLE, n=10) with or without RF, were analyzed for SARS-CoV-2 antibodies using 17 commercially available lateral flow assays (LFA), two ELISA kits and one in-house developed IgG multiplex bead-based assay. Six LFA and the in-house validated IgG assay correctly produced negative results for all samples. However, the majority of assays (n=13), gave false positive signal for samples from patients with RA and SLE. This was most notable in samples from RF positive RA patients. No false positive samples were detected in any assay using samples from patients with MS. Poor specificity of commercial serological assays could possibly be, at least partly, due to interfering antibodies in samples from patients with chronic inflammatory diseases. For these patients, the risk of false positivity should be considered when interpreting results of the SARS-CoV-2 serological assays.

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.

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