Human papillomavirus ‘reflex’ testing as a screening method in cases of minor cytological abnormalities

Abstract

The aim was to evaluate human papillomavirus (HPV) 'reflex genotyping' in cases of minor cytological abnormalities detected in the gynaecological screening programme in Stockholm, Sweden. Liquid-based cytology samples showing minor cytological abnormalities were analysed using HPV genotyping (Linear Array, Roche diagnostics). Colposcopically directed cervical biopsies were obtained and the HPV test results were correlated with the histological results. In all, 63% (70/112) of the samples were high-risk (HR) HPV (HR-HPV) positive. A statistically significant correlation was found between high-grade cervical lesions and HR-HPV (P ¼ 0.019), among which HPV 16, 18, and 31 were the most important. The negative predictive value of HR-HPV detection for histologically confirmed high-grade lesions was 100%. An age limit for HPV reflex testing may be motivated in cases of low-grade squamous intraepithelial neoplasia (LSIL), because of high HR-HPV prevalence among younger women. By using HPV reflex genotyping, additional extensive workup can safely be avoided in about 50% of all cases of atypical squamous cells of undetermined significance (ASCUS) and LSIL among women X30 years. This screening strategy could potentially reduce the total abnormal cytology-reporting rate in the Swedish screening programme by about 1% and provide more accurately directed follow-up, guided by cytological appearance and HPV test results.

Age-specific prevalence of HPV genotypes in cervical cytology samples with equivocal or low-grade lesions

“…These findings suggest that age 30, or even younger, is a suitable cutoff point for HR-HPV triage in LSIL, when consideration is given only to the prevalence argument. This is supported by a histological follow-up study of 112 of these cases by our group, in which 48% of women over 30 years with LSIL and 60% of women with ASCUS were HR-HPV negative and none of these had a histology result of CIN2 þ (Froberg et al, 2008) and could therefore have avoided additional investigation. This implies that the number of follow-ups after low-grade or ambiguous cytology results could be cut in half, at least in women over 30 years of age.…”

“…From September 2005 through to September 2006, 4204 LBC samples from a population-based screening programme were obtained from seven maternity health centres in Southern Stockholm as previously reported (Froberg et al, 2008). Later in 2006, an additional 2024 samples were collected.…”

“…The question whether HPV testing may improve the Swedish screening programme is under discussion, especially in the light of the potential to identify precancerous lesions among ASCUS and LSIL and to distinguish these from reactive or degenerative lesions. According to our previous study, complementing liquid-based cytology (LBC) with an HPV test in cases of ASCUS and LSIL may lead to a 40% decrease in the number of these women referred for further investigation (Froberg et al, 2008). Human papillomavirus prevalence is high among younger women with LSIL, which is why an age limit for triage HPV testing is being sought.…”

Comparison of the cobas Human Papillomavirus (HPV) Test with the Hybrid Capture 2 and Linear Array HPV DNA Tests

“…The cobas HPV test (cobas; Roche Molecular Systems, Pleasanton, CA) was recently approved by the U.S. Food and Drug Administration (FDA) and identifies HPV16 and HPV18 separately as well as detecting a pool of 11 HR-HPV genotypes (HPV31,) and also HPV66. The test has been validated in some initial studies (1, 13), and we sought to further add to the literature by assessing the interassay agreement between cobas and two other well-validated HPV DNA assays using samples collected in specimen transport medium (STM) (Qiagen, Gaithersburg, MD).Specifically, we compared cobas to (i) Linear Array (LA) (Roche Molecular Systems), an HPV genotyping assay that, while not approved by the FDA, is widely used for research (4,5,11,14) and is CE marked for use in Europe, and (ii) the FDA-approved Hybrid Capture 2 assay (HC2) (Qiagen Corporation, Gaithersburg, MD), which targets the 13 HR-HPV genotypes and cross-reacts with HPV66 (as well as a few other possibly carcinogenic or low-risk types) (2). …”

Slumbering mucosal immune response in the cervix of human papillomavirus DNA-positive and -negative women

“…DNA was extracted with the MagNA Pure LC robot and analyzed with the Linear Array HPV genotyping test (both procedures by Roche). Details of this method have been published (33). The Linear Array test is a PCR-and probe hybridization-based genotyping assay covering 37 HPV types including 12 high-risk HPV types (16, 18, Table II.…”