6th ESACP Congress, Heidelberg, April 7-11, 1999

A118
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

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.