Test Catalog

Test Id : SHPV

Human Papillomavirus (HPV) DNA Detection with Genotyping, High-Risk Types by PCR, SurePath, Varies

Useful For
Suggests clinical disorders or settings where the test may be helpful

Detection of high-risk (HR) genotypes associated with the development of cervical cancer

 

An aid in triaging women with abnormal Pap smear results

 

Individual genotyping of human papillomavirus (HPV)-16 and/or HPV-18, if present

 

This test is not recommended for evaluation of suspected sexual abuse.

Method Name
A short description of the method used to perform the test

Real-Time Polymerase Chain Reaction (PCR)

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.

Yes

Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

HPV with Genotyping, PCR, Surepath

Aliases
Lists additional common names for a test, as an aid in searching

High Risk HPV

HPV (Human Papillomavirus) PCR

Human Papillomavirus (HPV) Genotyping

Specimen Type
Describes the specimen type validated for testing

Varies

Necessary Information

Specimen source, collection date, and patient identifiers are required.

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Supplies: HPV SurePath Transport Tube 13 mL (T710)

Specimen Type: Cervical (endocervical or ectocervical) or vaginal

Specimen Volume: 1.5 mL

Collection Instructions:

1. Aliquot a minimum of 1 mL SurePath specimen into SurePath HPV aliquot tube.

2. Bag specimens individually as they have a tendency to leak during transport.

3. Place labels on the vial and on the bag.

Forms

If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the Testing Laboratory

1 mL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

SurePath enriched cell pellet Reject

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Varies Ambient (preferred) 14 days
Refrigerated 14 days

Useful For
Suggests clinical disorders or settings where the test may be helpful

Detection of high-risk (HR) genotypes associated with the development of cervical cancer

 

An aid in triaging women with abnormal Pap smear results

 

Individual genotyping of human papillomavirus (HPV)-16 and/or HPV-18, if present

 

This test is not recommended for evaluation of suspected sexual abuse.

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

Persistent infection with human papillomavirus (HPV) is the principal cause of cervical cancer. The presence of HPV has been implicated in more than 99% of cervical cancers worldwide, including both cervical squamous cell carcinoma and cervical adenocarcinoma. Before the development of invasive cancer, HPV infects the squamous mucosa cells and/or the glandular cells of the endocervix, leading to clonal expansion and morphologic changes. While the HPV-infected cells are restricted to their normal anatomic location, these changes are classified as cervical intraepithelial neoplasia (CIN). The severity of the morphologic changes and the degree to which those changes resemble the morphology of an invasive carcinoma are used to "grade" CIN. In general, high-grade CIN more closely resembles invasive carcinoma morphologically. HPV can also infect other mucosal cells in the anogenital region, such as the vaginal mucosa, leading to the development of HPV-associated intraepithelial neoplasia as well as invasive carcinoma not involving the cervix itself, although this is less common.

 

HPV is a small, nonenveloped, double-stranded DNA virus, with a genome of approximately 8000 nucleotides. There are more than 118 different types of HPV and approximately 40 different HPVs can infect the human anogenital mucosa. Only a very small percentage of patients who are exposed to HPV will develop CIN. Of those patients, only a small percentage will progress to invasive cervical cancer. Sexually transmitted infection with HPV is extremely common, with estimates of up to 75% of all women being exposed to HPV at some point. However, almost all infected women will mount an effective immune response and clear the infection within 2 years without any long-term health consequences. Both high-risk HPV genotypes (especially HPV-16 and 18), as well as persistent HPV infection (eg, an infection that is not cleared by the patient's immune system over time), are associated with an increased chance of progressing to high-grade CIN and invasive cancer.

 

Data suggest that certain HPV genotypes types (eg, HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) are considered high-risk (HR) for the development of cervical cancer and its precursor lesions. Furthermore, HPV types 16 and 18 have been regarded as the genotypes most closely associated with progression to cervical cancer. HPV-16 is the most carcinogenic, and is associated with approximately 60% of all cervical cancers, while HPV-18 accounts for approximately 10% to 15% of cervical cancers.(1-3)

 

In developed countries with cervical cancer screening programs, the Pap smear has been used since the mid-1950s as the primary tool to morphologically detect CIN, the precursor to cervical cancer. Pap smear screening has decreased death rates due to cervical cancer dramatically, since in many cases CIN can be treated and eliminated (eg, by local excision) before it progresses to invasive carcinoma. Although Pap smears and other liquid-based cytology methods have many advantages, they also have limitations: they require subjective interpretation by a highly trained cytopathologist and misinterpretation can occur, morphologic changes that resemble HIV-associated CIN can be caused by other conditions (eg, inflammation), and Pap smear does not sample every cell within the cervix/anogenital region potentially leading to falsely negative results. Perhaps most importantly, Pap smear does not differentiate between HPV genotypes that are high or low risk for progression to cervical cancer and it does not detect very early infections, which may lack a morphological phenotype.

Nucleic acid (DNA) testing by polymerase chain reaction has become a standard, noninvasive method for determining the presence of a cervical HPV infection. Proper implementation of nucleic acid testing for HPV may 1) increase the sensitivity of cervical cancer screening programs by detecting high-risk lesions earlier in women 30 years and older with normal cytology and 2) reduce the need for unnecessary colposcopy and treatment in patients 21 and older with cytology results showing atypical squamous cells of undetermined significance (ASC-US).

 

Recently, data suggest that individual genotyping for HPV types 16 and 18 can assist in determining appropriate follow-up testing and triaging women at risk for progression to cervical cancer. Studies have shown that the absolute risk of CIN-2 or worse in HPV-16 and/or HPV-18 positive women is 11.4% (95% CI, 8.4%-14.8%) compared with 6.1% (95% CI, 4.9%-7.2%) of women positive for "other" HR-HPV genotypes and 0.8% (95% CI, 0.3%-1.5%) in HR-HPV negative women.(4) Based in part on these data, the American Society for Colposcopy and Cervical Pathology (ASCCP) now recommends that HPV 16/18 genotyping be performed on women who are positive for HR-HPV, but negative by routine cytology/Pap smear. Women who are found to be positive for HPV-16 and/or -18 may be referred to colposcopy, while women who are negative for genotypes 16 and/or 18 may have repeat cytology and HR HPV testing in 12 months.(1)

Reference Values
Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.

Negative for HPV genotypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68

Interpretation
Provides information to assist in interpretation of the test results

A positive result indicates the presence of human papillomavirus (HPV) DNA due to 1 or more of the following genotypes: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68.

 

A negative result indicates the absence of HPV DNA of the targeted genotypes.

 

For patients with atypical squamous cells of undetermined significance (ASC-US) Pap smear result and who are positive for high-risk (HR) HPV, consider referral for colposcopy, if clinically indicated.

 

For women aged 30 years and older with a negative Pap smear result but who are positive for HPV-16 and/or HPV-18, consider referral for colposcopy, if clinically indicated.

 

For women aged 30 years and older with a negative Pap smear, positive HR HPV test result, but who are negative for HPV-16 and HPV-18, consider repeat testing by both cytology and a HR HPV test in 12 months.

Cautions
Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

The cobas human papillomavirus (HPV) test is FDA-approved for cervical/endocervical samples collected in PreservCyt (ThinPrep) media. Other sample types (eg, vaginal) collected in media, such as SurePath, are not considered FDA-approved sources; however, verification studies have been completed in compliance with CLIA-regulations by Mayo Clinic Laboratories.

 

Prolonged storage (>14 days) of clinical samples in SurePath media may impact the detection of high-risk (HR) HPV, especially if the amount of nucleic acid present in the sample is initially at a low concentration. Therefore, samples should be submitted for testing as soon as possible following collection.

 

The cobas HPV test detects DNA of the high-risk types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68. This test does not detect DNA of low-risk HPV types (eg, 6, 11, 42, 43, 44), which are not associated with invasive cervical cancer and its precursor lesions. Low-risk HPV types are associated with noninvasive genital warts and laryngeal papillomatosis.

 

Prevalence of HPV infection in a population may affect performance. Positive predictive values decrease when testing populations with low prevalence or individuals with no risk of infection.

 

Infection with HPV is not an indicator of cytologic high grade intraepithelial lesion (HSIL) or high-grade cervical intraepithelial neoplasia (CIN), nor does it indicate that a high-grade intraepithelial lesion (eg, HSIL or CIN2-3) or cancer will develop. Most women infected with 1 or more HR HPV types do not develop CIN2-3 or cancer.

 

A negative HR HPV result does not exclude the possibility of a patient developing a high-grade intraepithelial lesion (eg, HSIL or CIN2-3) or cancer in the future.

Supportive Data

To assess the accuracy of the Roche cobas human papillomavirus (HPV) test using cervical/endocervical and vaginal samples collected in SurePath media, a combination of spiking and comparison testing was performed. For spiking studies, 30 analyte-negative clinical samples (cervical/endocervical or vaginal matrix in SurePath media) were spiked with AcroMetrix HPV positive genotype controls (type 68 [n=10], type 16 [n=10], type 18 [n=10]) at 1 dilution above the limit of detection (LOD). The results are summarized in Table 1 below:

 

Table 1. Verification of accuracy for the Roche cobas HPV test using spiked cervical/endocervical samples in SurePath media.

Source

Positive

Negative

Agreement

HPV Genotype 68

10/10

0

100%

HPV Genotype 16

10/10

0

100%

HPV Genotype 18

10/10

0

100%

 

In addition to the spiking studies described above, clinical samples (n=26) collected in SurePath media and initially tested by the Roche cobas HPV assay at an outside laboratory were tested at Mayo Clinic Laboratories (MCL) in a blinded fashion. The results are summarized in Table 2 below:

 

Table 2. Comparison of SurePath samples tested by Roche cobas HPV at an outside laboratory and MCL.

 

Roche cobas 4800 - outside laboratory

Roche cobas 4800 - MCL

 

Positive

Negative

Total

Positive

14

14

14

Negative

2

10

12

Total

16

10

26

 

Agreement: 92.3% (74.7-99.0%)

 

Reference Range:

Cervical/endocervical samples (n=27) and vaginal samples (n=22) collected in SurePath media for routine Pap smear screening were tested by the Roche cobas HPV assay.

 

All 49 samples (100%) had negative Pap results and negative Roche cobas HPV 4800 results.

 

The reference range for the Roche cobas HPV test is negative.

 

LOD (Analytical Sensitivity):

To assess the analytical sensitivity of the Roche cobas HPV test, pools of cervical/endocervical/vaginal specimens in SurePath media were created. Pools were spiked at a high starting concentration using each of the 3 AcroMetrix HPV Genotype controls (cell lines infected with HPV genotypes 16, 18, or 68). Serial dilutions were made into analyte-negative sample containing cells to achieve dilution of the analyte to the point of extinction. At least 6 replicates of each dilution were tested, including the panel member that was 1 dilution below the predicted LOD. The LOD was established as the highest dilution in which 6 of 6 replicates were positive.

 

The LOD of the Roche cobas HPV genotype 16, 18, and "Other" high-risk HPV infected cells in SurePath media was determined to be 50 cells/mL, 1250 cells/mL, and 250 cells/mL, respectively.

 

Analytical Specificity:

A full specificity panel has been tested by the manufacturer that included bacteria, fungi and viruses, including those commonly found in the female urogenital tract. Also, several HPV types classified as low or undetermined risk were tested with the cobas HPV test to assess analytical specificity. Results indicated that none of these organisms interfered with detection of HPV 31, HPV16, and HPV18 or produced false-positive results from specimens negative for high-risk HPV

 

Specimen Stability:

The stability of SurePath samples (endocervical/cervical and vaginal) at ambient (18-24 degrees C) was assessed using spiking studies and clinical samples. These results are summarized in Tables 3 and 4 below:

 

Table 3. Negative SurePath samples spiked with AcroMetrix positive genotype controls (68, 16 or 18). Samples were held at ambient temperature for 14 days.

 

Type 68-

5000 cells/mL

Type 16-

5000 cells/mL

Type 18-

5000 cells/mL

Day

Crossing point

Crossing point

Crossing point

0

31.6

31.8

36.7

7

32.5

31.4

36.6

14

34.2

31.9

39.2

AVERAGE

32.3

31.7

37.5

% CV

4.03

0.83

3.93

 

In addition to the spiking studies described above, clinical samples collected in SurePath media at an outside laboratory were held at ambient temperature over a period of 14 days and tested by the Roche cobas HPV assay.

 

Table 4. Positive SurePath pooled patient material collected at an outside laboratory were held at ambient temperature and tested over 14 days.

 

Patient genotype

"Other HR HPV"

Patient Genotype

HPV-16

Patient Genotype

HPV-18

Day

Crossing point

Crossing point

Crossing point

0

37.2

27.5

29.6

7

36.4

28.7

30.4

14

37.4

28.9

29.8

AVERAGE

37.0

28.4

29.9

% CV

1.43

2.66

1.39

Clinical Reference
Recommendations for in-depth reading of a clinical nature

1. Saslow D, Solomon D, Lawson HW, et al: American Cancer Society, American Society for Colposcopy and Cervical Pathology, and American Society for Clinical Pathology screening guidelines for the prevention and early detection of cervical cancer. J Low Genit Tract Dis. 2012;16(3):175-204. doi: 10.1097/LGT.0b013e31824ca9d5.  

2. Walboomers JM, Jacobs MV, Manos MM, et al: Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol. 1999;189:12-19. doi: 10.1002/(SICI)1096-9896(199909)189:1

3. de Sanjose S, Quint WG, Alemany L, et al: Human papillomavirus genotype attribution in invasive cervical cancer: a retrospective cross-sectional worldwide study. Lancet Oncol. 2010;11:1048-1056. doi: 10.1016/S1470-2045(10)70230-8.

4. Wright TC Jr, Stoler MH, Sharma A, et al: Evaluation of HPV-16 and HPV-18 genotyping for the triage of women with high-risk HPV positive, cytology-negative results. Am J Clin Pathol. 2011 Oct;136(4):578-586. doi: 10.1309/AJCPTUS5EXAS6DKZ.

5. Gilbert L, Oates E, Ratnam S: Stability of cervical specimens in SurePath medium for HPV testing with the Roche cobas 4800. J Clin Microbiol. 2013 Oct;51(10):3412-3414. doi: 10.1128/JCM.01391-13

Method Description
Describes how the test is performed and provides a method-specific reference

The cobas human papillomavirus (HPV) test targets and detects nucleic acid from the L1 region of the HPV genome using real-time polymerase chain reaction (PCR) technology. The cobas HPV test is used for the in vitro qualitative detection of 14 high-risk HPV types commonly associated with cervical cancer. The assay is able to specifically assess for the presence or absence of HPV genotypes 16 and 18 while concurrently detecting the remaining 12 high-risk types (31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68). The cobas HPV test is used in conjunction with the cobas 4800 System. The cobas 4800 System comprises the cobas x 480 instrument and cobas z 480 analyzer that fully automates the cobas HPV from sample extraction through amplification, detection, and data reduction.(Instruction manual and package insert: cobas HPV test. Roche Diagnostics. version 05641268001-20EN. 03/2021)

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information

No

Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Monday through Friday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

3 to 6 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

1 week

Performing Laboratory Location
Indicates the location of the laboratory that performs the test

Rochester

Fees
Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

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  • Clients without access to Test Prices can contact Customer Service 24 hours a day, seven days a week.
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Test Classification
Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

This test has been modified from the manufacturer's instructions. Its performance characteristics were determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information
Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.

87624

G0476 (if appropriate)

LOINC® Information

Test Id Test Order Name Order LOINC Value
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.
SHPV HPV with Genotyping, PCR, Surepath 77378-8
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
SS018 Specimen Source 31208-2
36003 HPV High Risk type 16, PCR 61372-9
36004 HPV High Risk type 18, PCR 61373-7
36005 HPV other High Risk types, PCR 77375-4

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

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Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports