Slides were prepared on a ThinPrep processor (T5000, Hologic Inc) using special ThinPrep slides (Hologic, Inc) and stained using CINtec PLUS test kits within 48 hours and processed on BenchMark ULTRA system (Roche Diagnostics) per manufacturer’s instructions.
The CINtec PLUS slides were initially evaluated independently by one of two experienced cytotechnologists who were trained to read these slides. Smears were determined to be positive if at least one cervical epithelial cell showed both a brownish cytoplasmic immunostaining for p16 and a red nuclear immunostaining for Ki-67 regardless of cellular morphology. If the dual staining was not observed, the smear was considered negative. Smears were deemed unsatisfactory if they did not contain an adequate number of cells ( 4 cells per field with a minimum of 10 fields with a 40x objective). All slides were independently reviewed by a study pathologist trained to read CINtec PLUS slides, and the results recorded using the same criteria. Discrepant slides were either internally reviewed by another reader and reconciled or adjudicated independently by an external expert.
The CINtec PLUS slides were initially evaluated independently by one of two experienced cytotechnologists who were trained to read these slides. Smears were determined to be positive if at least one cervical epithelial cell showed both a brownish cytoplasmic immunostaining for p16 and a red nuclear immunostaining for Ki-67 regardless of cellular morphology. If the dual staining was not observed, the smear was considered negative. Smears were deemed unsatisfactory if they did not contain an adequate number of cells ( 4 cells per field with a minimum of 10 fields with a 40x objective). All slides were independently reviewed by a study pathologist trained to read CINtec PLUS slides, and the results recorded using the same criteria. Discrepant slides were either internally reviewed by another reader and reconciled or adjudicated independently by an external expert.