No evidence for interference of h&e staining in DNA testing: usefulness of DNA extraction from H&E-stained archival tissue sections.
Although histochemical staining has been believed to inhibit the DNA amplification reaction, no previous study has systematically evaluated the influence of histochemical staining on downstream molecular assays. To evaluate an influence of H&E staining on DNA testing, we isolated DNA from 10 unstained, 10 hematoxylin-stained, 10 eosin-stained, and 10 H&E-stained tissue sections (ie, 4 groups), from each of 5 colon cancers. Among the 4 groups, we did not observe any significant or appreciable difference in DNA fragmentation by agarose gel electrophoresis, in DNA amplification by real-time polymerase chain reaction (PCR), in microsatellite PCR fragment analyses, or in a PCR-pyrosequencing assay. As a proof-of-principle study, we successfully performed microsatellite instability analysis and sequencing of KRAS and BRAF on more than 1,300 colorectal cancers using DNA extracted from H&E-stained tissue sections. Our data provide no evidence for an interfering effect of H&E staining on DNA testing, suggesting that DNA from H&E-stained sections can be effectively used for routine DNA testing.
Center for Molecular Oncologic Pathology, Dana-Farber Cancer Institute, Harvard Medical School, 450 Brookline Ave, Room JF-208E, Boston, MA 02215, USA.
Teppei
T
Morikawa
Kaori
K
Shima
Aya
A
Kuchiba
Mai
M
Yamauchi
Noriko
N
Tanaka
Yu
Y
Imamura
Xiaoyun
X
Liao
Zhi Rong
ZR
Qian
Mohan
M
Brahmandam
Janina A
JA
Longtine
Neal I
NI
Lindeman
Charles S
CS
Fuchs
Shuji
S
Ogino
D003106:Colon / Q000473:pathology; D015179:Colorectal Neoplasms / Q000235:genetics / Q000473:pathology; D004396:Coloring Agents; D004273:DNA, Neoplasm / Q000032:analysis*; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin; D006801:Humans; D016133:Polymerase Chain Reaction; D012007:Rectum / Q000473:pathology; D013194:Staining and Labeling
D004396:Coloring Agents; D004273:DNA, Neoplasm; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin