Application of Polymerase Chain Reaction to Analysis of Antigen Receptor Rearrangements to Support Endoscopic Diagnosis of Canine Alimentary Lymphoma

Kaneko, Naoki; Yamamoto, Yoshihiko; Wada, Yuko; Miyama, Takako Shimokawa; Hiraoka, Hiroko; Itamoto, Kazuhito; Mizuno, Takuya; Nakaichi, Munekazu; Takahashi, Tomoko; Watari, Toshihiro; Okuda, Masaru

doi:10.1292/jvms.71.555

Cited by 27 publications

(28 citation statements)

References 17 publications

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“…In contrast to previous studies, we also analyzed both not applicable native and denatured-reannealed products (heteroduplex analysis). 31 Denaturation and reannealing do not affect bands amplified from truly clonal cells but will produce a smear for pseudoclonal cell populations. Failure to eliminate pseudoclones may result in loss of specificity of up to 10%.…”

Section: Discussionmentioning

confidence: 99%

Distinguishing Intestinal Lymphoma From Inflammatory Bowel Disease in Canine Duodenal Endoscopic Biopsy Samples

et al. 2014

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Inflammatory bowel disease (IBD) and intestinal lymphoma are intestinal disorders in dogs, both causing similar chronic digestive signs, although with a different prognosis and different treatment requirements. Differentiation between these 2 conditions is based on histopathologic evaluation of intestinal biopsies. However, an accurate diagnosis is often difficult based on histology alone, especially when only endoscopic biopsies are available to differentiate IBD from enteropathy-associated T-cell lymphoma (EATL) type 2, a small cell lymphoma. The purpose of this study was to evaluate the utility of histopathology; immunohistochemistry (IHC) for CD3, CD20, and Ki-67; and polymerase chain reaction (PCR) for antigen receptor rearrangement (T-cell clonality) in the differential diagnosis of severe IBD vs intestinal lymphoma. Endoscopic biopsies from 32 dogs with severe IBD or intestinal lymphoma were evaluated. The original diagnosis was based on microscopic examination of hematoxylin and eosin (HE)-stained sections alone followed by a second evaluation using morphology in association with IHC for CD3 and CD20 and a third evaluation using PCR for clonality. Our results show that, in contrast to feline intestinal lymphomas, 6 of 8 canine small intestinal lymphomas were EATL type 1 (large cell) lymphomas. EATL type 2 was uncommon. Regardless, in dogs, intraepithelial lymphocytes were not an important diagnostic feature to differentiate IBD from EATL as confirmed by PCR. EATL type 1 had a significantly higher Ki-67 index than did EATL type 2 or IBD cases. Based on the results of this study, a stepwise diagnostic approach using histology as the first step, followed by immunophenotyping and determining the Ki67 index and finally PCR for clonality, improves the accuracy of distinguishing intestinal lymphoma from IBD in dogs.

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Section: Discussionmentioning

confidence: 99%

Distinguishing Intestinal Lymphoma From Inflammatory Bowel Disease in Canine Duodenal Endoscopic Biopsy Samples

et al. 2014

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“…PCR testing for clonal rearrangement of lymphocyte antigen was performed for 8 dogs in the Shiba group, and TCR gene clonal rearrangement was detected in 2 dogs. As the sensitivity and specificity of PCR testing are reportedly questionable to some extent for detecting alimentary lymphoma [9,17], there is some possibility that these were false-positive results. Unfortunately, necropsy could not be conducted on the 2 dogs, and we could not completely exclude the possibility of lymphoma.…”

Section: Discussionmentioning

confidence: 99%

A Retrospective Study in 21 Shiba Dogs with Chronic Enteropathy

Ohmi

Ohno

Uchida

et al. 2011

The Journal of Veterinary Medical Science

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ABSTRACT. We retrospectively studied the clinical and laboratory features and outcomes of chronic enteropathy in Shiba dogs. Among 99 dogs with chronic enteropathy, 21 Shiba dogs (21%) were included in the study (odds ratio, 7.14). No significant differences were seen in signalment, clinical signs, symptoms or laboratory profiles between the Shiba and non-Shiba groups. Severe histopathological lesions in the duodenum were a common finding in the Shiba group. The median overall duration of survival in the Shiba group was 74 days, while that of the dogs in the non-Shiba group could not be determined because more than half of the cases remained alive at the end of this study. The difference between the groups was statistically significant (P<0.0001). The 6-month and 1-year survival rates for the Shiba group were 46% and 31%, respectively. Conversely, the 6-month, 1-year and 3-year survival rates for the non-Shiba group were 83%, 74% and 67%. The results obtained here demonstrated that the Shiba dog is predisposed to chronic enteropathy and shows severe duodenum lesions and poor outcomes, indicating a breed-specific disease.KEY WORDS: breed-specific enteropathy, chronic enteropathy, Shiba.

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“…2,13 It is reportedly useful for the diagnosis of canine alimentary lymphoma. 4,7 This technique is highly sensitive but tends to produce false-positive results, a phenomenon termed pseudoclonality, possibly because a single copy is not amplified during PCR because of the immunologic diversity of the target genes: immunoglobulin heavy chain (IgH) and T-cell receptor gamma (TCRc). To avoid diagnostic errors, it is necessary to determine the reliability of the results of PARR.…”

Section: <!?Show "Fnote_aff1"$^!"content-markup(/author-grp[1]/aff|/author-grp[1]/dept-list)>mentioning

confidence: 99%

Heteroduplex Polymerase Chain Reaction is Essential for Canine Receptor Rearrangement Analysis

Takanosu

Tadika

Kobayashi³

2010

J VET Diagn Invest

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Abstract. Polymerase chain reaction (PCR) for antigen receptor rearrangement is a sensitive technique for detecting lymphocyte-proliferative disorders, but it tends to produce false-positive results, a phenomenon termed pseudoclonality. Heteroduplex analysis, which is useful to distinguish clonal reactions from pseudoclonal ones in dogs, can be applied to avoid misdiagnosis and determine the reliability of results. In the current study, PCR for antigen receptor rearrangement was used to identify clonal proliferation of lymphocytes in duodenal and lymphoid tissue from dogs presenting with chronic vomiting and enlarged peripheral lymph nodes typical of multicentric lymphoma, and the test results were verified with heteroduplex analysis. In the case of almost all of the duodenal samples, even without a histologic diagnosis of lymphoma, a distinct band similar to that observed in the case of lymphoma was obtained for both B-and T-cell clonality. All of the bands obtained from the nonneoplastic duodenum disappeared following heteroduplex analysis of the PCR product, whereas the distinct bands from the lymphoma remained. In the lymph node samples, the pseudoclonal bands that disappeared in the heteroduplex analysis were detected mainly in B cells. In conclusion, heteroduplex analysis with PCR for antigen receptor rearrangement is a suitable tool for diagnosing canine lymphoma and decreasing the possibility of misdiagnosis of pseudoclonality.

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Application of Polymerase Chain Reaction to Analysis of Antigen Receptor Rearrangements to Support Endoscopic Diagnosis of Canine Alimentary Lymphoma

Cited by 27 publications

References 17 publications

Distinguishing Intestinal Lymphoma From Inflammatory Bowel Disease in Canine Duodenal Endoscopic Biopsy Samples

Distinguishing Intestinal Lymphoma From Inflammatory Bowel Disease in Canine Duodenal Endoscopic Biopsy Samples

A Retrospective Study in 21 Shiba Dogs with Chronic Enteropathy

Heteroduplex Polymerase Chain Reaction is Essential for Canine Receptor Rearrangement Analysis

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