Influence of histochemical and immunohistochemical stains on polymerase chain reaction.

Author(s): Murase T, Inagaki H, Eimoto T

Publication: Mod Pathol, 2000, Vol. 13, Page 147-51

PubMed ID: 10697271 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this paper was to determine the effects of prior histological or IHC staining and antigen retrieval method on PCR success using template DNA from FFPE lymph nodes.

   Summary of Findings:

   An average of 5 additional PCR cycles were necessary to amplify a 110 bp region of beta-globin from hematoxylin and May-Grunwald stained sections compared to eosin Y or methyl green stained or unstained sections. There was no effect of prior immunostaining without antigen retrieval when the detection was with DAB or fuchsin. A slight but non-significant increase in required PCR cycles was noted for immunostained sections where staining was with DAB-cobalt. The use of antigen retrieval significantly inhibited PCR success, but specimens unmasked by trypsin digestion were amplifiable. Heat-induced antigen retrieval using a microwave or pressure cooker resulted in 2 and 5 specimens, respectively, failing to amplify after 60 cycles.

   Biospecimens

   • Tissue - Lymph Node

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic

   Platform:

   Analyte	Technology Platform
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Antigen retrieval	Trypsin in PBS at 37 degrees C Microwave 10 min in citrate buffer Pressure cooker
   PCR Specific	Type of tissue stain	Mayers hematoxylin Eosin Y Methyl green May-Grunwald solution None DAB DAB-cobalt Fuchsin
   PCR Specific	Targeted nucleic acid	Beta-globin

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