Processing of long-stored archival cervical smears for human papillomavirus detection by the polymerase chain reaction

Abstract

Summary The efficiency of a freeze-thaw method, a proteinase K/Tween 20 lysis method and a guanidinium isothiocyanate/silica beads method for DNA extraction from fixed and Papanicolaou-stained cells from the cervical cancer cell line Siha was measured by P-globin polymerase chain reaction (PCR). The GTC/silica beads method, which appeared superior, revealed a human papillomavirus (HPV) general primer-mediated

A systematic review of the role of human papilloma virus (HPV) testing within a cervical screening programme: summary and conclusions

“…However, many of the most relevant studies are just beginning to reach the literature, and most of the large studies related to screening are still ongoing with at most only preliminary reports available. Currently, two consensus primer PCR systemsthe MY09/11 and the GP5+/6+ pairs -and the second-generation Hybrid Capture system (HC-II) with high risk probes would seem to be the methods of choice (Manos et al, 1989;de Roda Husman et al, 1995;Peyton et al, 1998). These three methods all have high absolute sensitivity for detecting oncogenic viruses and have the potential for automation.…”

“…With modern tests, over 95% of all cancers are HPV positive (Bosch et al, 1995;Walboomers et al, 1999;Cuzick et al, 2000) and 75-95% of high-grade CIN lesions are associated with a positive HPV test on exfoliated cells (de Roda Husman et al, 1995;Herrington et al, 1995;Burger et al, 1996;Clavel et al, 1999;Cuzick et al, 1999). These and other comparative studies have shown that HPV testing has a greater sensitivity for CIN II/III than cytology.…”

“…This variability reflects a number of factors, including age, extent of sexual exposure, previous disease, and type of assay used. Overall the positivity rate of tests for high-risk HPV types in women not known to have CIN was 13% ranging from 10% for PCR using GP5/6 primers (de Roda Husman et al, 1995;Kruger-Kjaer et al, 1998) to 20% using Hybrid Capture II (Clavel et al, 1999;Cuzick et al, 1999;Hill et al, 1999). Rates are about half these levels in women aged over 35 years.…”

Human papillomavirus testing in cervical cancer screening

“…Although Grainge et al (2005) found a significantly higher HPV positivity among cases compared to controls, a substantial proportion of women tested negative for HPVs in baseline smears less than 4 years (67%), and 4 -13 years (74%), before the diagnosis of CIN3/cervical cancer. Unfortunately, such use of long-stored archived cervical smears is prone to the danger of false-negative HPV findings (De Roda, 1995) and crosscontamination (Chua and Hjerpe, 1995).It is important to bear in mind, however, that the detection of HPV at a single point in time (as in case -control studies) or over a few years of follow-up (seldom more than 5 years in available cohort studies, Schiffman et al, 2005) does not allow an accurate estimate of a woman's lifelong risk of HPV infection nor of cervical cancer. Transiently detectable HPV infection is generally regarded as harmless, but the long-term prognosis of an HPV-positive finding is not fully understood.…”

“…Although Grainge et al (2005) found a significantly higher HPV positivity among cases compared to controls, a substantial proportion of women tested negative for HPVs in baseline smears less than 4 years (67%), and 4 -13 years (74%), before the diagnosis of CIN3/cervical cancer. Unfortunately, such use of long-stored archived cervical smears is prone to the danger of false-negative HPV findings (De Roda, 1995) and crosscontamination (Chua and Hjerpe, 1995).…”