Cervista hpv hr test

A liquid-based cervical cytology smear was taken with a cervical brush and immediately transferred to 20 mL of Thin-Prep PreservCyt Solution (Hologic, Beijing). The ThinPrep-2000 automatic cell detector was used for programmed processing. Cervical cytology diagnosis is classified into five levels according to the 2014 Bethesda System (TBS) criteria [25 (link)]. Abnormal TCT results are as follows: atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion (ASC-H), high-grade squamous intraepithelial lesion (HSIL) and atypical glandular epithelial cells (AGC) within 3 months.
The Cervista HPV HR test (Hologic, Beijing) was an in vitro diagnostic test for the qualitative detection of 14 hrHPV DNA types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68). When hrHPV detection was positive, we used the Cervista HPV 16/18 test (Hologic, Beijing) to assess the presence or absence of hrHPV16 and 18.

The Cervista HPV HR test (Hologic Inc., Madison, WI, USA) is a qualitative test detecting 14 hrHPV types (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68) [13] (link), [14] . The assay uses three separate oligonucleotide mixtures; Mix 1 (A5/A6 pool) contains probes for HPV 51, 56 and 66; mix 2 (A7 pool) probes for HPV 18, 39, 45, 59 and 68, and mix 3 (A9 pool) probes for HPV 16, 31, 33, 35, 52 and 58. In these three mixes, oligonucleotides for the human histone 2 gene (HIST2H2BE) are also present as an internal control for the presence of sufficient genomic DNA [14] . A signal to noise value (sample signal measured against signal from a No Target Control) is generated for each of the three mixes and is referred to as HPV Fold-Over-Zero (FOZ). The HPV FOZ ratio is calculated by dividing the highest FOZ value from any one of the three reaction mixtures by the lowest HPV FOZ value of the three mixtures. If the HPV FOZ ratio is equal to or greater than 1.525, the sample is considered positive for hrHPV [14] . Samples with mixed HPV infections might result in positive signals of similar intensity in two or three reaction wells. Therefore, if the HPV FOZ ratio is lower than 1.525, but the HPV FOZ values in all three mixes are larger than the second cut-off value at 1.93 (default setting), the sample is considered positive for hrHPV in the Cervista HPV HR test [14] .

All ThinPrep® LBC samples sent to Manchester were analysed by staff trained and certified by Hologic (Hologic, Bedford, MA). HPV testing of LBC samples was carried out using the FDA-approved Cervista HPV HR test [23 (link)] in conjunction with the Cervista MTA (Hologic) automated platform according to the manufacturer’s instructions. Prior to and after analysis, all LBC samples were stored at +4°C in a monitored refrigerator.