Abstract: Methods and reagents for detecting high-risk human papilloma virus (HPV) DNA types in cells on a Pap smear that indicates the patient is at higher risk for cancer are described. The method uses full-length DNA probes to HPV types 16, 18, 31, 33, 35, and 51 in a particular proportion to hybridize to and detect the viral DNA in situ. The method differentiates high-risk from low-risk HPV DNA in cells that indicates the patient's risk for cancer. The in situ hybridization is detected by brightfield microscopy.

Abstract: Improvements to the in situ polymerase chain reaction (PCR), a process of in vitro enzymatic amplification of specific nucleic acid sequences within the cells where they originate, can be achieved by changing the way that the enzymatic reaction is started. Reaction initiation is delayed until the start of PCR thermal cycling, either by withholding a subset of PCR reagents from the cellular preparation until the preparation has been heated to 50.degree. C. to 80.degree. C., immediately before thermal cycling is begun, or by adding to the PCR reagents a single-stranded DNA binding protein which blocks reaction at temperatures below about 50.degree. C. If the in situ PCR is performed on cellular preparations already attached to a microscope slide, thermal cycling also is facilitated by use of a thermal cycler sample block or compartment designed optimally to hold the microscope slide and any vapor barrier covering the slide.