Analytical characterization of the APTIMA HPV Assay.
BACKGROUND
Human papillomavirus (HPV) testing has improved the sensitivity for the detection of cervical pre-cancer and cancer as compared to Pap testing. Several HPV tests are commercially available and most target the DNA from 13 or 14 high-risk HPV types. The APTIMA HPV Assay however, detects HPV E6/E7 mRNA from 14 high-risk types of HPV: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68.


OBJECTIVE
To determine the analytical performance characteristics of the APTIMA HPV Assay.


METHODS
Analytical sensitivity, analytical specificity, reproducibility, and the effect of potentially interfering substances was determined for the APTIMA HPV Assay on both the DTS (semi-automated) and TIGRIS DTS (fully automated) systems.


RESULTS
The 95% detection limit for both systems was between 17 and 488 copies/reaction, depending on the HPV type. The assay did not cross-react with normal flora and opportunistic organisms that may be found in cervical samples, or low-risk HPV types. Spermicides, anti-fungal and anti-itch medications, whole blood, glacial acetic acid, and most lubricants did not interfere with assay performance. Those lubricants containing polyquaternium 15 did interfere with assay performance. Inter-instrument, inter-operator, inter-lot, and inter-run signal variability were <10% for >99% of the data. Intra-run variability was <15%, except for those samples with concentrations at or below the 95% detection limit of the assay.


CONCLUSIONS
Based upon the analytical sensitivity, analytical specificity, and low variability, the APTIMA HPV Assay showed excellent performance and robustness.
Dockter
Janel
J
Gen-Probe Incorporated, San Diego, California 92121, USA. janeld@gen-probe.com
Schroder
Astrid
A
Eaton
Barbara
B
Wang
Ann
A
Sikhamsay
Nathan
N
Morales
Liezel
L
Giachetti
Cristina
C
D001331:Automation; D002584:Cervix Uteri / Q000821:virology; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D027383:Papillomaviridae / Q000472:pathogenicity*; D030361:Papillomavirus Infections / Q000821:virology*; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis*
D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic