Skip to main content
. 2018 Sep 18;4(4):213–226. doi: 10.1002/cjp2.111

Table 1.

Commercially available assays and reagents for HPV detection

Assay types* Method/commercial kits Technical details/interpretation/evaluation Web information
p16 IHC CINtec® Histology , § (Roche mtm Lab, Heidelberg, Germany) Qualitative IHC test using mouse monoclonal anti‐p16 antibody clone E6H4. Used in light microscopic assessment of p16INK4a protein in FFPE tissues https://usdiagnostics.roche.com/en/document/CINtec-Histology-Interpretation-Guide.pdf
CINtec® Histology § (Ventana Medical Systems, Tucson, AZ, USA) http://www.ventana.com/product/1743?type=2202
mAb clone JC8 (Santa Cruz Biotechnology Inc., Dallas, TX, USA); mAb clone G175‐405 (BD Biosciences, San Jose, CA, USA) Other anti‐p16 antibodies for IHC (other than E6H4 clone) http://www.nordiqc.org/downloads/assessments/26_59.pdf
In situ hybridization for HPV DNA GenPoint™ HPV DNA Probes Cocktail (Y1443, Dako, CA, USA) Probes contain HPV genomic clones in the form of double‐stranded fragments of 500 bp or less (biotinylated and unlabeled) and multiple biotinylated oligonucleotides from 25 to 40 bases in length.
Staining procedure uses Dako GenPoint™ Detection System (code K0620), a nonfluorescent chromogen, and observed under light microscope. React with HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68 on FFPE tissues and/or cells by ISH. Indicate for HPV genomic integration
https://www.agilent.com/cs/library/packageinsert%20/public/128074001.PDF
GenPoint™ HPV type 16/18 biotinylated DNA probes (Y1412, Dako, CA, USA) Use labeled DNA probes that specifically bind to a consensual or type‐specific HR‐HPV DNA sequences and detect amplified signals. React with HPV types 16, 18, on FFPE tissues and/or cells by ISH http://www.finels.com/product/up_files/Y1412.pdf
Bond™ Ready‐to‐Use ISH HPV Probe (subtypes 16, 18, 31, 33, 51) (Leica Microsystems, IL, USA) Qualitative identification of the HPV DNA in FFPE tissue by ISH using the automated BOND system (includes Leica BOND‐MAX system and Leica BOND‐III system). This probe binds to the five high‐risk HPV subtypes, 16, 18, 31, 33 and 51 https://www.leicabiosystems.com/fileadmin/img_uploads/novocastra_reagents/Novocastra_datasheets/pb0829.pdf
INFORM HPV III Family 16 Probe (B)§ (Ventana Medical Systems, AZ, USA) Contains a cocktail of HPV genomic probes in a formamide‐based diluent. The intended targets are the common high‐risk HPV genotypes found to be associated with neoplasia. The probe cocktail has demonstrated affinity to the following genotypes: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, and 66 http://www.uclad.com/newsletters/HPV_ISH_Tissue-Probe-Interpretation_Guide.pdf
In situ hybridization for HPV RNA RNAscope® HPV Test (Advanced Cell Diagnostics, CA, USA) RNAscope® HPV Biomarker Detection Reagents and its proprietary ‘double Z’ oligonucleotide probes specific for each subtype HPV E6/E7 mRNA enable high specificity detection of viral transcripts in routine FFPE tumor biopsies. Labeled probes contain chromogenic enzyme or fluorophore signal generating one punctate dot per RNA target. Hybridization of only three Z probe pairs is sufficient to obtain a detectable chromogenic signal by a brightfield microscope. Detect HPV 16, 18, 31, 33, 35, 52, and 58 https://acdbio.com/science/applications/disease-areas/hpv-related-cancer
HPV 16 mRNA Probe (Ventana Medical System, Tucson, AZ, USA) DNP hapten labeled probe designed to bind to the HPV 16 E6/E7 mRNA transcript. The probe has affinities to the HPV 16 genotype with no known cross reactivity to other common HPV genotypes http://www.ventana.com/product/1830?type=2340
PCR and liquid‐phase assays Digene® Hybrid Capture 2 (HC2) High‐Risk HPV DNA Test§ (Qiagen, Gaithersburg, MD, USA) In vitro nucleic acid hybridization assay with signal amplification using antibodies and chemiluminescence for qualitative detection of 13 high‐risk types of HPV DNA. Probes are based on GP5+/GP6 primer set and detect HPV L1 open reading frame. Detects high‐risk HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68. Cannot be used to determine specific HPV type present https://www.qiagen.com/us/shop/detection-solutions/human-pathogens/digene-hc2-high-risk-%20hpv-dna-test/#orderinginformation
Digene® HPV Genotyping PS Test For use with Digene HC2 High‐Risk HPV DNA test for qualitative genotyping of HPV 16, 18, and 45 https://www.qiagen.com/gb/shop/detection-solutions/human-pathogens/digene-hpv-%20genotyping-ps-test/#orderinginformation
Digene® HC2 HPV DNA Test , § In vitro microplate assay based on an RNA probe cocktail and signal‐amplified nucleic acid hybridization that uses chemiluminescence for the qualitative detection of 13 high‐risk (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68) and 5 low‐risk types of HPV DNA (6, 11, 42, 43, 44). Can differentiate between two HPV DNA groups, low‐risk and high‐risk types https://www.qiagen.com/us/shop/detection-solutions/human-pathogens/digene-hc2-hpv-dna-%20test/#productdetails
LINEAR ARRAY® HPV Genotyping Test (Roche Diagnostics, Indianapolis, IN, USA) Based on PGMY09/11 primers set, detect HPV L1 open reading frame. A qualitative test that detects 37 high‐and low‐risk HPV genotypes https://molecular.roche.com/assays/linear-array-hpv-genotyping-test
INNO‐LiPA HPV Genotyping extra II(20T) (Fujirebio Europe, Ghent, Belgium) SPF10 Plus primers set provide high test sensitivity due to the precision of the short 65‐base pair PCR product. Permits simultaneous detection of multiple genotypes in a single sample. One probe line for one genotype, ready‐to‐use master mix for RT‐PCR application, and automation. Individually detect 32 HPV genotypes http://search.cosmobio.co.jp/cosmo_search_p/search_gate2/docs/IGT_/81534.20160810.pdf
Cobas® HPV Test , § (Roche Molecular Diagnostics, Pleasanton, CA, USA) Uses automated specimen preparation to extract DNA. Amplifies HPV DNA by PCR followed by nucleic acid hybridization for automated real‐time detection of 14 high‐risk HPV types in a single analysis. Specimens are limited to cells collected in PreservCyt® Solution, cobas® PCR Cell Collection Media and SurePath® Preservative Fluid. Test concurrently identifies 14 high‐risk HPV types (HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) and can also differentiate between HPV 16 and HPV 18 https://diagnostics.roche.com/global/en/products/params/cobas-hpv.html
The Abbott RealTime High Risk (HR) HPV assay (Abbott Park, IL, USA) Qualitative in vitro PCR assay that utilizes homogeneous target amplification and detection technology for detection of high‐risk HPV DNA in cells collected in liquid cytology media. Detect 14 high‐risk HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68 and to partially genotype 16, 18 from other 12 high‐risk genotypes https://www.molecular.abbott/int/en/products/%20infectious-disease/realtime-high-risk-hpv
Cervista® HPV HR Test , § (Hologic Inc., Bedford, MA, USA) Uses Invader™ chemistry, a signal amplification method for detection of specific nucleic acid sequences. This method uses two types of isothermal reactions: a primary reaction that occurs on the targeted DNA sequence and a secondary reaction that produces a fluorescent signal. Qualitative detection of DNA from 14 high‐risk HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68. Cannot determine the specific HPV type https://www.hologic.com/sites/default/files/package-insert/15-3100_105_01.pdf
Cervista™ HPV 16/18 Test , § Detects HPV DNA 16, 18 https://www.hologic.com/sites/default/files/package-insert/15-3101_102_01.pdf
Aptima® HPV assay , § (Hologic Inc., Bedford, CA, USA) Nucleic acid amplification involving 3 steps that take place in a single tube: target capture; target amplification of RNA by transcription‐mediated amplification (TMA); and detection of amplification products (amplicons) by hybridization protection assay (HPA). Assay incorporates an internal control to monitor nucleic acid capture, amplification, and detection, as well as operator or instrument error. Detects E6/E7 viral mRNA from 14 high‐risk HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68 https://www.hologic.com/sites/default/files/2018-02/AW-12821_002_01.pdf
Onclarity™ HPV assay , § (BD Diagnostics Systems, Baltimore, MD, USA) Amplified DNA test for the qualitative detection of high risk types of human HPV. Using BD Onclarity HPV Cervical Brush Collection Kit, BD SurePathT™ Preservative Fluid, andPreservCyt® Solution. Performed with the BD Viper™ LT System. Detects all high‐risk HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68), and provides the capability for genotyping of six high risk types (HPV 16, 18, 31, 45, 51, and 52) http://www.puls-norge.no/media/285557/pakningsvedlegg-hpv.pdf
*

Most assays are tested and validated in cervical cancer. The assays are listed from the most commonly use pathological tests to the more technical challenged or advanced methodologies.

Technical details only describe the main technical principles of the methods or commercial kits. Each methods or kits may have different protocol details and reagents.

CE marked for HPV detection for cervical cancer in Europe. CE marking is a certification mark that indicates conformity with health, safety, and environmental protection standards for products sold within the European Economic Area (EEA). CE Marking indicates a product's compliance with the applicable EU regulations and enables the commercialization of products in 32 European countries.

§

FDA approved for HPV detection for cervical cancer in United States.

Additional antibodies available for p16 IHC were tested and used in clinical pathology laboratory. The information are summerized in the weblink to NordiQC website.