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Molecular diagnosis of Ewing's sarcoma/primitive neuroectodermal tumor in formalin-fixed paraffin-embedded tissues by RT-PCR and fluorescence in situ hybridization.
Diagn Mol Pathol. 2005 Mar; 14(1):23-8.DM

Abstract

Recent studies have suggested that formalin-fixed paraffin-embedded (FFPE) tissues can be used for molecular analyses by fluorescence in situ hybridization (FISH) and RT-PCR. We analyzed 18 cases of ES/PNET for the t(11;22)(q24;q12) and t(21;22)(q22;q12) fusion transcripts by RT-PCR and analyzed for EWS translocation by interphase FISH with a dual color fusion probe to compare these two approaches directly. RT-PCR detected 13 (72%) EWS-FLI-1 fusions (type I=10, type II=3) and 2 (11%) EWS-ERG fusions. Three cases could not be evaluated because the housekeeping gene phosphoglycerate kinase (internal mRNA control) was not amplified. FISH was diagnostic in 15 of 18 cases (83%). There were three discordant cases between RT-PCR and FISH (concordance of 83%). Using a combination of RT-PCR and FISH, the results were complementary. One advantage of RT-PCR analysis was that subtypes of EWS translocation could be determined specifically (type I, type II and ERG). These findings indicate that because of the difficulties and limitations associated with the molecular analysis of FFPE tissues, a combination of RT-PCR and FISH may be a better approach to enhance the sensitivity and accuracy of detecting ES/PNET translocations in FFPE tissues with suboptimally preserved nucleic acids.

Authors+Show Affiliations

Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota 55905, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

15714060

Citation

Qian, Xiang, et al. "Molecular Diagnosis of Ewing's Sarcoma/primitive Neuroectodermal Tumor in Formalin-fixed Paraffin-embedded Tissues By RT-PCR and Fluorescence in Situ Hybridization." Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B, vol. 14, no. 1, 2005, pp. 23-8.
Qian X, Jin L, Shearer BM, et al. Molecular diagnosis of Ewing's sarcoma/primitive neuroectodermal tumor in formalin-fixed paraffin-embedded tissues by RT-PCR and fluorescence in situ hybridization. Diagn Mol Pathol. 2005;14(1):23-8.
Qian, X., Jin, L., Shearer, B. M., Ketterling, R. P., Jalal, S. M., & Lloyd, R. V. (2005). Molecular diagnosis of Ewing's sarcoma/primitive neuroectodermal tumor in formalin-fixed paraffin-embedded tissues by RT-PCR and fluorescence in situ hybridization. Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B, 14(1), 23-8.
Qian X, et al. Molecular Diagnosis of Ewing's Sarcoma/primitive Neuroectodermal Tumor in Formalin-fixed Paraffin-embedded Tissues By RT-PCR and Fluorescence in Situ Hybridization. Diagn Mol Pathol. 2005;14(1):23-8. PubMed PMID: 15714060.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular diagnosis of Ewing's sarcoma/primitive neuroectodermal tumor in formalin-fixed paraffin-embedded tissues by RT-PCR and fluorescence in situ hybridization. AU - Qian,Xiang, AU - Jin,Long, AU - Shearer,Brandon M, AU - Ketterling,Rhett P, AU - Jalal,Syed M, AU - Lloyd,Ricardo V, PY - 2005/2/17/pubmed PY - 2005/7/1/medline PY - 2005/2/17/entrez SP - 23 EP - 8 JF - Diagnostic molecular pathology : the American journal of surgical pathology, part B JO - Diagn Mol Pathol VL - 14 IS - 1 N2 - Recent studies have suggested that formalin-fixed paraffin-embedded (FFPE) tissues can be used for molecular analyses by fluorescence in situ hybridization (FISH) and RT-PCR. We analyzed 18 cases of ES/PNET for the t(11;22)(q24;q12) and t(21;22)(q22;q12) fusion transcripts by RT-PCR and analyzed for EWS translocation by interphase FISH with a dual color fusion probe to compare these two approaches directly. RT-PCR detected 13 (72%) EWS-FLI-1 fusions (type I=10, type II=3) and 2 (11%) EWS-ERG fusions. Three cases could not be evaluated because the housekeeping gene phosphoglycerate kinase (internal mRNA control) was not amplified. FISH was diagnostic in 15 of 18 cases (83%). There were three discordant cases between RT-PCR and FISH (concordance of 83%). Using a combination of RT-PCR and FISH, the results were complementary. One advantage of RT-PCR analysis was that subtypes of EWS translocation could be determined specifically (type I, type II and ERG). These findings indicate that because of the difficulties and limitations associated with the molecular analysis of FFPE tissues, a combination of RT-PCR and FISH may be a better approach to enhance the sensitivity and accuracy of detecting ES/PNET translocations in FFPE tissues with suboptimally preserved nucleic acids. SN - 1052-9551 UR - https://www.unboundmedicine.com/medline/citation/15714060/Molecular_diagnosis_of_Ewing's_sarcoma/primitive_neuroectodermal_tumor_in_formalin_fixed_paraffin_embedded_tissues_by_RT_PCR_and_fluorescence_in_situ_hybridization_ L2 - https://doi.org/10.1097/01.pdm.0000140192.27878.97 DB - PRIME DP - Unbound Medicine ER -