Use of Formalin-Fixed Paraffin-Embedded Tissue and Single-Strand Conformation Polymorphism Analysis for Polymerase Chain Reaction of Antigen Receptor Rearrangements in Dogs

Kaneko, Naoki; Tanimoto, Takashi; Morimoto, Masahiro; Hayashi, Toshiharu; Miyama, Takako Shimokawa; Hiraoka, Hiroko; Itamoto, Kazuhito; Une, Satoshi; Mizuno, Takuya; Okuda, Masaru

doi:10.1292/jvms.71.535

Cited by 10 publications

(8 citation statements)

References 13 publications

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“…These samples were originated from years of 1997, 2006, 2011 and 2012. It has been reported that not long‐term paraffin‐embedding but the formalin fixation time for more than 1 week can impede PCR . Since the fixation time at our institute is limited to 24–48 hours, we attribute the negative results of PCR with Cmu primers to small tissue sample sizes, which may have increased formalin permeability into tissue samples and thus intensified negative effects by formalin fixation on PCR …”

Section: Discussionmentioning

confidence: 98%

Detection of monoclonality in intestinal lymphoma with polymerase chain reaction for antigen receptor gene rearrangement analysis to differentiate from enteritis in dogs

Ohmura

Leipig

Schöpper

et al. 2015

Vet Comparative Oncology

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The diagnosis of canine intestinal lymphoma by morphological examination is challenging, especially when endoscopic tissue specimens are used. The utility of detection of antigen receptor gene rearrangement by polymerase chain reaction (PARR) in canine lymphoma has been well established, but its usefulness to distinguish enteritis and intestinal lymphoma remains unclear. In this retrospective study we assessed clonality of 29 primary canine intestinal lymphoma, 14 enteritis and 15 healthy control cases by PARR analysis, using formalin-fixed, paraffin-embedded full-thickness tissue specimens. We could detect monoclonal rearrangements in 22 of 29 canine intestinal lymphomas [76%; 95% confidence interval (CI) 56-90%] and polyclonal rearrangements in all of the enteritis and healthy control cases (100%; CI 88-100%). We revealed a predominance of T-cell phenotype compared to B-cell phenotype (85%; CI 65-96% and 15%; CI 4-35%, respectively). We showed that PARR analysis contributes to differentiation of canine intestinal lymphoma from enteritis and to phenotyping of lymphomas.

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Section: Discussionmentioning

confidence: 98%

Detection of monoclonality in intestinal lymphoma with polymerase chain reaction for antigen receptor gene rearrangement analysis to differentiate from enteritis in dogs

Ohmura

Leipig

Schöpper

et al. 2015

Vet Comparative Oncology

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“…Removal of paraffin from the tissue is the most critical step for successful extraction as undissolved paraffin which leads to poor sample quality and inhibition of PCR amplification [12]. Naoki Kaneko et al, demonstrated that formalin fixation time and percentage play an important role in getting good quality of amplifiable DNA for PCR analysis [13]. There is another factor, quantification of DNA which also influence the PCR results from FFPE tissue DNA.…”

Section: Discussionmentioning

confidence: 99%

Quantification of DNA Extracted from Formalin Fixed Paraffin-Embeded Tissue Comparison of Three Techniques: Effect on PCR Efficiency

Kumar

Panigrahi

Suryavanshi

et al. 2016

JCDR

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“…To avoid diagnostic errors, it is necessary to determine the reliability of the results of PARR. Single-strand conformation polymorphism is useful for verifying the results of conventional PARR, 6 but the procedure involved is relatively complicated and time consuming. On the other hand, heteroduplex analysis is reportedly useful to distinguish clonal reactions from pseudoclonal ones in cats 9 and dogs 11 .…”

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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Use of Formalin-Fixed Paraffin-Embedded Tissue and Single-Strand Conformation Polymorphism Analysis for Polymerase Chain Reaction of Antigen Receptor Rearrangements in Dogs

Cited by 10 publications

References 13 publications

Detection of monoclonality in intestinal lymphoma with polymerase chain reaction for antigen receptor gene rearrangement analysis to differentiate from enteritis in dogs

Detection of monoclonality in intestinal lymphoma with polymerase chain reaction for antigen receptor gene rearrangement analysis to differentiate from enteritis in dogs

Quantification of DNA Extracted from Formalin Fixed Paraffin-Embeded Tissue Comparison of Three Techniques: Effect on PCR Efficiency

Heteroduplex Polymerase Chain Reaction is Essential for Canine Receptor Rearrangement Analysis

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