![]() |
6th ESACP Congress, Heidelberg, April 7-11, 1999 |
A118
We have developed a protocol for processing cytological material of the
uterine cervix that allows cytomorphological evaluation and additional
molecular and cytochemical analyses, such as immunochemistry (IC), in situ
hybridization (ISH) and human papilloma virus (HPV) PCR typing. We have
evaluated several fixation protocols and optimized the logistics. In short,
cervical brush material is smeared unto microscope slides and rest material
is collected in Unifix primary fixative by the gynaecologist and send to
the cytology laboratory. Within 72 hrs the cells are centrifuged and
resuspended in agar. Solid agar cell blocks are subsequently embedded in
paraffin (Agarcyto) and 5 µm sections are used for further molecular analysis.
Cytomorphology of Agarcyto preparations closely resemble cytological smears
but show histological features as well allowing to identify tissue patterns
not recognizable in smears. We show that IC detection of Ki-67, p53, bcl-2,
ISH for centromere of chromosome 1, and HPV typing by PCR is feasible on
serial sections from of one cytological sample. We are currently using the
Agarcyto preparation technique to prospectively draw a molecular inventory
of cervical premalignancies.
AGARCYTO CELL BLOCK PREPARATION OF CERVICAL BRUSH MATERIAL FOR
MULTI-PARAMETER MOLECULAR ANALYSIS.
Robben H, Wienk S, Kerstens H, Gemmink A, Macville M,
Poddighe P 1, Boonstra H 2, de Wilde P, Hanselaar A
Institute of Pathology, University Hospital Nijmegen, 1) Clinical Genetics
Center, University Hospital Utrecht, 2) Dept. of Gynaecology and Obsterics,
University Hospital Nijmegen, the Netherlands