Identification of neoplastic cells in blood using the Sysmex XT‐2000iV: a preliminary step in the diagnosis of canine leukemia

Gelain, Maria Elena; Rossi, G.; Giori, Luca; Comazzi, S.; Paltrinieri, Saverio

doi:10.1111/j.1939-165x.2010.00214.x

Cited by 12 publications

(9 citation statements)

References 18 publications

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“…In addition, the overall relative rat bone marrow differential counts obtained in the studies here are comparable to ranges established over the past 10 years using the antibody‐specific flow cytometric method. Also, gating of blasts within the bone marrow 5‐part differential count was consistent with a previous report of blast identification using the Sysmex platform in peripheral blood of dogs with leukemia . Most importantly, once established, gating can be consistently used without further adjustment, although a certain expertise and training with the Sysmex instrument is required.…”

Section: Discussionsupporting

confidence: 87%

Validation of Sysmex XT‐2000iV generated quantitative bone marrow differential counts in untreated Wistar rats

Criswell

Bock

Wildeboer

et al. 2014

Veterinary Clinical Pathol

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Background: Preclinical drug trials frequently require assessment of bone marrow toxicity in animals to evaluate hematopoietic safety. Since the gold standard, cytologic evaluation, is time consuming and requires highly trained individuals, automated methods remain intriguing. Objective: The Sysmex XT-2000iV hematology analyzer allows userdeveloped customizable gating. This study was conducted to validate the gating of bone marrow cell populations in Sysmex cytograms from untreated rats. Methods: B-and T-lymphocytes and myeloid cells were experimentally depleted from Charles River Wistar Han IGS (CRL: WI [Han]) rat whole bone marrow suspension using a magnetic cell sorting (MACS) method. The positively and negatively selected populations were used to verify select gates within the Sysmex cytogram. Intra-and inter-animal precision, comparability between right and left femur, as well as agreement with microscopic myelograms based on 500 counted cells, were determined. Results: Intra-sample precision and right-to-left femur comparability confirmed that gating was reproducible and stable. In 50 tested rats, myeloid to erythroid ratios (M:E) were 1.32 AE 0.33 in males and 1.38 AE 0.29 in females by Sysmex compared to 1.36 AE 0.32 in males and 1.42 AE 0.32 in females by microscopic evaluations. Bland-Altman differences between methods was ≤ AE0.35 units for M:E, ≤ 5.4% for maturing myeloid cells, ≤ 3.4% for proliferating myeloid cells, ≤ 6.0% for maturing myeloid cells, ≤ 3.4% for proliferating myeloid cells, and ≤ 4.1% for lymphocytes. Conclusions: In untreated control Charles River Wistar Han IGS (CRL: WI [Han]) rats, the Sysmex XT-2000iV produced an automated M:E and 5-part differential count equivalent to microscopic differential counts.

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Section: Discussionsupporting

confidence: 87%

Validation of Sysmex XT‐2000iV generated quantitative bone marrow differential counts in untreated Wistar rats

Criswell

Bock

Wildeboer

et al. 2014

Veterinary Clinical Pathol

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“…Although flow cytometry can be rapid and informative and may provide a very high likelihood of lymphoma and even phenotype, it is unable to provide a diagnosis of specific lymphoma subtype. 9,18,25 The practice of lymphoma diagnosis in human medicine is solidly based on histology, and this practice needs to be followed in veterinary oncology as well. Currently, most lymphoma therapy in animals is largely palliative, suggesting that collaboration is needed with oncologists to test specific protocols on single subtypes of lymphoma with a goal of therapy that is closer to curative.…”

Section: Discussionmentioning

confidence: 99%

Canine Lymphomas

et al. 2013

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Canine lymphoma is the neoplasm most often treated by chemotherapy, yet there are few data to correlate response to therapy with its different subtypes. This study is based on biopsy specimens from 992 dogs for which lymphoma was the clinical diagnosis. All cases were phenotyped by immunohistochemistry for CD3 and CD79alpha. Cases with histiocytic proliferation were evaluated immunohistochemically for CD18. Clonality was verified in 12 cases by polymerase chain reaction (PCR). Survival (event time) data and complete survival information (cause of death or time to last follow-up) were available on 456 dogs. Additional covariate information when available included size, age, sex, phenotype, stage and grade of lymphoma, mitotic index, and treatment protocol. Because of the many subtypes of B-and T-cell lymphoma, the cases were grouped into 7 diagnostic categories: (1) benign hyperplasia; (2) low-grade B-cell; (3) high-grade B-and T-cell; (4) low-grade T-cell; (5) centroblastic large B-cell of all mitotic grades (subdivided by clinical stage); (6) immunoblastic large B-cell of all mitotic grades, and (7) high-grade peripheral T-cell. Grouping was determined by histological grade (based on mitotic rate/400Â field, with low-grade 0-5, intermediate 6-10, and high-grade >10) and stage for survival function estimation. No association with survival was found for size (based on breed of dog) or sex. All diagnostic categories of indolent or low-grade type had low mitotic rates, whereas those with clinically high grades had high mitotic rates. The diagnostic category with the most cases was centroblastic large B-cell lymphoma. Compared with dogs in this largest represented group of lymphomas, dogs with high-grade lymphomas had significantly higher mortality rates, and dogs with low-grade T-cell lymphomas had significantly lower mortality rates. Treatments for high-, intermediate-, and low-grade lymphomas were divided into 4 groups: absence of treatment, chemotherapy with or without hydroxydaunorubicin, and only prednisone. Dogs with low-grade T-cell (T-zone) lymphomas had the longest median survival (622 days), whereas the shortest median survival was in dogs with T-cell high-grade (peripheral T-cell) subtype (162 days). The dogs with centroblastic large B-cell lymphomas had a median survival of 127 days with low stage, 221 days with intermediate stage, and 215 days with advanced stage. Dogs with T-zone lymphoma were probably diagnosed in later stages of disease because of the lack of signs associated with progression. As with human lymphomas, a histological diagnosis with immunophenotyping is a minimal requirement for diagnosis of a specific subtype.

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“…The Sysmex XT‐2000iV is a recent hematology analyzer that combines impedance and optical methods and for which specific veterinary software has been developed. The analyzer has been used in veterinary medical research and has been validated for use in dogs . Reference intervals (RIs) aid in the interpretation of laboratory results by characterizing normal fluctuations observed in a healthy population, and have only recently been established for dogs using the Sysmex analyzer .…”

Section: Introductionmentioning

confidence: 99%

Establishment of canine hematology reference intervals for theSysmexXT‐2000iV hematology analyzer using a blood donor database

Serra¹,

Freeman²,

Campora³

et al. 2012

Veterinary Clinical Pathol

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Identification of neoplastic cells in blood using the Sysmex XT‐2000iV: a preliminary step in the diagnosis of canine leukemia

Abstract: Using a novel gating strategy the Sysmex XT-2000iV may be used effectively to screen canine blood for hematopoietic neoplasia.

Cited by 12 publications

References 18 publications

Validation of Sysmex XT‐2000iV generated quantitative bone marrow differential counts in untreated Wistar rats

Validation of Sysmex XT‐2000iV generated quantitative bone marrow differential counts in untreated Wistar rats

Canine Lymphomas

Establishment of canine hematology reference intervals for theSysmexXT‐2000iV hematology analyzer using a blood donor database

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