Comparison of GP5+/6+-PCR and SPF₁₀-Line Blot Assays for Detection of High-Risk Human Papillomavirus in Samples from Women with Normal Cytology Results Who Develop Grade 3 Cervical Intraepithelial Neoplasia

Abstract

Using a case control approach, we performed a two-way comparison study between GP5؉/6؉-PCR and HPV SPF 10 -Line Blot 25 (SPF 10 ) assays for detection of 14 types of high-risk human papillomavirus (hrHPV) in samples from women with normal cytology results who had or developed grade 3 cervical intraepithelial neoplasia (CIN 3). Samples were pooled from two cohorts, i.e., women participating in population-based screening and women attending a gynecological outpatient clinic. Cases (n ‫؍‬ 45) were women with histologically confirmed CIN 3 diagnosed within a median follow-up time of 2.7 (range, 0.2 to 7.9) years. Control samples were from women (n ‫؍‬ 264) who had developed CIN 1 lesions at maximum (median follow-up at 5. Based on the fact that a persistent infection with high-risk human papillomavirus (hrHPV) is the necessary cause of cervical cancer (4, 20), hrHPV testing has been recognized as a potentially valuable tool not only for the triage of women with borderline cytomorphological abnormalities but also for primary cervical screening, either in conjunction with cytology testing or not. Support for this comes from various clinical studies (1,6,9,10,16,26). It should be realized, however, that hrHPV tests might detect both transient and persistent hrHPV infections, of which only the latter represent a condition that ultimately may result in the development of grade 3 cervical intraepithelial neoplasia (CIN 3) lesions and cervical cancer (ՆCIN 3) and therefore should be considered as clinically relevant. We previously hypothesized that various hrHPV tests, although similarly efficient in detecting persistent infections, may differ considerably in the extent with which transient, clinically irrelevant hrHPV infections are detected (28).A clinically valuable hrHPV test should perform in such a way that the number of transient infections detected is as low as possible to ensure an optimal balance between clinical sensitivity and specificity for ՆCIN 3. This is important to minimize the number of unnecessary follow-up procedures, particularly for women with normal cytology results. Based on data from longitudinal clinical studies involving large cohorts, so far only the GP5ϩ/6ϩ-PCR and the commercially available hrHPV hybrid capture 2 (hc2) assays have been proven to reach such an optimal balance and therefore can be considered clinically validated (2,5,7,9,16,17,21,24). In a previous two-way comparison study, we showed that these assays, which both detect hrHPV types as a pool, perform equally well for the detection of ՆCIN 3 in a population-based cervical screening setting (13).Besides GP5ϩ/6ϩ-PCR and hc2, the consensus HPV SPF 10 -Line Blot 25 linear probe assay (SPF 10 -LiPA version 1) PCR assay (15) (referred to herein as SPF 10 ) is a commonly used HPV detection assay, particularly in epidemiological studies and vaccination trials (12). However, this method has not yet

High-Risk Human Papillomavirus in Esophageal Squamous Cell Carcinoma

“…In addition, although there is no universally accepted "gold standard" primer set for detecting HPV DNA in formalin-fixed, paraffin-embedded tissues, the GP5+/GP6+ primers we used have been shown to be highly sensitive for detecting HPV DNA in exfoliated cervical cells (48), and have been used previously for detecting HPV DNA in formalin-fixed tissues (49). Taken together, these features give confidence that the prevalence estimate and reported associations are unlikely to be biased by the selection of participants or the collection of data.…”

Dry Storage and Transport of a Cervicovaginal Self-Sample by Use of the Evalyn Brush, Providing Reliable Human Papillomavirus Detection Combined with Comfort for Women

“…Each DNA isolation and PCR test run contained HPV-positive and -negative controls. All self-collected and physician-obtained samples were tested for HPV with both the analytically sensitive SPF 10 -PCR system (29,30) and the clinically validated GP5ϩ/6ϩ-PCR-based test (25,35).…”

Technical Issues Associated With Estimating the Prevalence of HPV-52 in Cervical Intraepithelial Neoplasia

“…The generic primer pairs MY09/11 and GP5+/6+ similarly contain mismatches to the HPV-52 L1 target sequence and are less sensitive for the detection of HPV-52 at lower viral DNA copy number. 52,55,56 GP5+/6+ has also been reported to have very low sensitivity for HPV-52 in a Chinese cohort of cervical cancer specimens due to a particular sequence variation in the target sequence that is overrepresented outside of Europe. 57 The PGMY09/11 consensus primer set was based on the MY09/11 generic primer pair and has improved detection of several genotypes, including HPV-52, due to a decreased number of primer mismatches.…”