Test for the detection and genotyping of High and Low HPV

High PapillomaStrip and High+Low PapillomaStrip are two tests based on the reverse hybridization principle, which allows the qualitative detection and genotyping of 19 and 37 subtypes (respectively) of Human Papillomavirus (HPV) in DNA samples extracted from cervico-uterine smears or biopsies, penis swabs or urine.

Human papillomavirus is the more common sexual transmitted disease (STD) worldwide. Of all the existing HPV genotypes, more than 40 can infect the anogenital tract and ⅓ of these are associated with cervical cancer and anal neoplasm. In fact, more than 90% of these cancers are caused by untreated HPV infections, resulting in the fourth most common cancer detected in women globally.

According to the WHO, there are three detection methods for the screening of precancerous lesions, including high-risk HPV testing. In comparison, this method is the most specific and it can detect the presence of the virus even before the appearance of the lesions, helping professionals to make an earlier and more accurate diagnosis of the disease.

High Papillomavirus test allows the qualitative and individual detection of up to 19 of the most common medium-high risk HPV genotypes in DNA samples from cervical smears or biopsies, penile smears and urine.

The High+Low PapillomaStrip test allows the qualitative and individual detection of 37 HPV genotypes (18 low-risk and 19 high-medium-risk) in DNA samples from cervical smears or biopsies, penile smears and urine in a single test.

Reliable

Detection of co-infections

37 subtypes in 1 test

Amplification of E6/E7 regions

Use of genotype-specific primers and probes

Our products for the detection and genotyping of High an Low HPV

Product	Technique	Sample	Catalogue No.
High+Low PapillomaStrip	Hybridization on Strip	DNA from cervical smears or biopsies, penile smears and urine	3.148.016.53.000
High+Low PapillomaStrip			3.148.048.53.000
High PapillomaStrip			3.146.016.53.000

Feature 1

Detection of 19 or 37 HPV subtypes in one test. This allows professionals to obtain more complete information for the diagnosis, as well as monitoring during the infection (they can verify whether or not the subtypes disappear) and after HPV vaccination.

Feature 2

Individual detection of all the subtypes. Non-clustered detection of the subtypes allows the identification of possible co-infections, as it provides more detailed information about the presence of those subtypes.

Feature 3

Use of genotype-specific primers and probes. This increases the specificity of the test and eliminates cross-reactions, making the result obtained more reliable.

Feature 4

Amplification of E6/E7 regions instead of L1. HPV PCR strategies targeting the E6/E7 region may be preferable to L1 due to the following reasons:

E6 and E7 are the oncogenic regions of HPV.
E6 and E7 exhibit strong sequence conservation even after infection, whereas E2 and L1 can be deleted.
The E6/E7 sequence differs between high and low-risk HPV types, thus providing diagnostic discrimination using PCR for testing.
E6 and E7 are conserved and stable regions. Those characteristics ensure its detection.

Publications

N.º	Title	Language
C1	Comparison of five automated CE/IVD labeled HPV DNA genotyping test systems	English
C2	Prevalence and genetic distribution of human Papyloma virus in women with cervical intraepithelial neoplasia	English
C3	Incidence of Chlamydia trachomatis infection in high risk human papillomavirus infected young women	English
C4	Preliminary evaluation of the High+Low PapillomaStrip Assay Colli-PeeTM collected UCM preserved urine	English
A1	Human papillonavirus infection in oral fluids of HIV-1-positive men: prevalence and risk factors	English
A3	Test Kitu Detekci a Genotypizaci HPV – High a Low Papilloma Strip 8 + 8 strips	Polish
A4	Identificación, Incidencia y Distribución de Genotipos del Virus del Papilloma Humano en una muestra de la población costarricense	Spanish
A5	Human papilloma virus genotyping in women with abnormal cytology (Ocena genotypów wirusów brodawczaka ludzkiego u kobiet z nieprawidlowa cytologia)	Polish
A7	In situ FoxP3+ and IL-10 over expression is associated with high grade anal lessions in HIV infected patients	English
A8	Human Papillomavirus Genotyping among Different Cervical Smears in Duhok/Irak	English
A9	Colli-Pee – Performance of a game-changing sampling device for HPV-based cervical cancer screening	English

Webinars

“New strategies for cervical cancer prevention”.

In this webinar, Dr. Paula Cortiñas, renowed gynecologist, talks about how to prevent cervical cancer and its direct link to HPV infections.

The recorder webinar is available on demand. If you missed it, feel free to request it!

information

Frequently Asked Questions

Why it is important to identify the type of HPV?

The anogenital HPVs are generally divided into two categories: those with a low oncogenic potential risk (Low-Risk Group) and those with a medium-high oncogenic potential risk (Medium-High-Risk Group). The high-risk HPVs are generally associated with high-grade precancerous lesions and invasive cancer (16 and 18 subtypes cause 70% of these lesions), while low-risk HPVs are frequently found in asymptomatic or benign conditions such as genital warts. These differences in the oncogenic potential of HPVs make necessary to identify the type of HPV present in the sample, which will allow much more targeted treatment at an earlier stage of the disease.

How is the dry swab or swab in PBS extracted?

When taking the swab, it is recommended to store it without any transport media. If PBS is added (maybe because other exams are made that require this PBS), then the sample should be stored frozen until the extraction. Otherwise, other microorganisms might grow, increasing DNA quantity and degrading the sample. Before the extraction, the dry swab is then resuspended in 500 µl of PBS in a 1,5 ml tube, mixing with vortex for 1 minute. After that, centrifugue the tube for 2 minutes, at 18.000 g. After discarding the supernatant, add 500 µl of PBS, and resuspend the pellet by vortex. Centrifugue the tube for 2 minutes, at 18.000 g. Discard the supernatant. Resuspend the pellet with 200 µl of PBS, and proceed as indicated in the protocol (20 µl of proteinase K + 20 µl buffer AL, etc). By doing that, interferent material will be eliminated, and the PCR will be “cleaner”.

How are sensitivities and specificities calculated for these products?

Generally, it is not possible to calculate a diagnostic sensitivity and specificity, but rather a concordance of sensitivity and specificity between 2 techniques, as neither is really the Gold Standard. In other words, only how one compares with the other can be said. On the other hand, the techniques compared do not detect exactly the same types. This means that, if one of the types not detected by one of the techniques is present, there will be a discordance. Not because of a false result, but because the test is not set up to detect it.

How should the sample be taken?

Use a dry, sterile cotton swab or brush for sampling. Be sure to take a sufficient amount of sample, but without bleeding from the lesion. Keep the swab in its tube, without using any preservative medium. Store it at 2/8 ºC if it is to be processed in less than a week or at – 20ºC if it is to be processed later.

Is it necessary for the patient to have lesions at the time of sampling?

No, the presence of lesions is not necessary. The diagnosis can be made before the appearance of lesions.

Should sampling be done exclusively by the gynecologist?

Self-collection kits have not been validated, in principle it is advisable that the collection is performed by a professional.

Why did I get a variable result when repeating the analysis?

One possible explanation is that the patient becomes infected and recovers between tests. If its a mild infection and the immune system is strong, the patient may overcome the infection and be re-infected with other types by the next test. There is also the possibility that the patient may have co-infections with several HPV types, but that all of them have a very low viral load. In these cases, it is possible that not all types will be amplified in each test, but that they will be amplified randomly. Since the types are not repeated, this does not seem likely, but cannot be ruled out. Another reason could be that there were contaminations in the different assays. It is unlikely to have occurred in several assays in a different way, but this should always be checked by placing blanks in all assays to ensure the veracity of the results.

Do repeated results for the same HPV type in different samples always imply co-infection?

Possibly contamination. Blanks should be tested: if they are correct, then the result is valid. This is not surprising because multiple infection in these cases is common. On the other hand, if the blank test comes back contaminated, then there is contamination. In this case, the reagents should be discarded and the cleaning protocol should be performed. No further samples should be tested until several valid blanks are obtained.

What happens if the cytology shows cellular changes associated with HPV infection, but the genotyping obtained with High HPV is negative?

It may be a type not present in the kit or at a low concentration.