The routine practical approach for diagnosing astrocytomas and oligodendrogliomas begins with perfoming IHC for ATRX and IDH1 R132H expression. Stepwise analysis of molecular parameters with initial IHC for ATRX and IDH1 R132H followed by 1p/19q analysis and then by IDH sequencing significantly reduces the number of molecular tests required for unequivocal diagnosis (Reus et al., 2015).

Characteristics of the 3 most important molecular groups of adult glioma:

Diffuse glioma with IDH mutation
and 1p/19q-codeletion

Diffuse glioma with
IDH mutation

Diffuse glioma without
IDH mutation

Biomarker

IDH1/2
1p/19q
ATRX
hTERT-Promotor

mutated
co-deleted
nuclear expression
mutated

mutated
intact
loss of nuclear expression
wildtype

wildtype
intact
nuclear expression
mutated

Typical histological finding and prognosis

Histology
WHO grading
Median Survival

oligodendroglial
II or III
>15 years

astrocytic
II or III (IV)
8-12 years

astrocytic
IV (II or III)
<2-3 years

Combined IHC on IDH1 R132H (clone H09) and ATRX (clone AX1) substitutes molecular testing.

Immunohistochemical staining (IHC) with anti-IDH1 R132H Antibody (clone H09) - dianova

Anti-IDH1 R132H
clone H09
#DIA-H09
0.5ml
1:20-1:50

Immunohistochemical staining (IHC) with anti-ATRX Antibody (clone AX1) - dianova

Anti-ATRX
clone AX1
#DIA-AX1
0.5ml
1:100-1:200

IDH1 R132H
The 2016 CNS WHO certification recommends IDH1 R132H IHC as a backbone for differential diagnosis of glioma. IDH1 R132H IHC is widely applied as a favorable prognostic marker.

ATRX
ATRX mutations in gliomas result in the loss of nuclear ATRX expression (right), which can be diagnosed by IHC analysis. Loss of ATRX expression is close to being mutually exclusive to 1p/19q co-deletion.

p53 and Ki67 are helpful accessory marker for classification of diffuse glioma.

Immunohistochemical staining (IHC) with anti-p53 Antibody (clone CC53) - dianova

Anti-p53
clone CC53
#DIA-530
0.5ml
1:100-1:200

Immunohistochemical staining (IHC) with anti-Ki-67/MIB1 Antibody (clone Ki-67P) - dianova

Anti-Ki67
clone Ki67P
#DIA-670
0.5ml
1:100-1:200

p53
p53 can be selected as a marker since there is evidence of relationships among p53, mutually exclusive to 1p/19q deletion, suggesting the usesfulness of ATRX and p53 IHC without 1p/19q analysis.

Ki-67
High Ki-67 is dominant in IDH wild type gliomas and low Ki-67 is associaterd with IDH1 mutation in primary glioblastomas. The mitotic index is significantly associated with outcome of IDH wild type tumors.

Reference:

Reuss DE et al. ATRX and IDH1-R132H immunohistochemistry with subsequent copy number analysis and IDH sequencing as a basis for an “integrated” diagnostic approach for adult astrocytoma, oligodendroglioma and glioblastoma. Acta Neuropathol. 129(1):133-146, 2015