HPV Low Risk (6/11) TaqMan PCR Kit
1 / 8Pages

3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 • (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Product Insert Product# TM32350 Intended Use Norgen’s HPV Low Risk (6/11) TaqMan PCR Kit is designed for the detection of 2 HPV low risk types (6/11) in a real-time PCR based on the use of TaqMan® technology. This kit is designed for research use only and not for use in diagnostic procedures. Background Information More than 70 types of human papillomavirus (HPV) have been identified, and are generally classified as high-risk or low-risk depending on their relationship or lack of relationship with cancer and high-grade cervical intraepithelial neoplasia (CIN 2-3). HPV viruses are predominantly sexually transmitted and high-risk HPV types are a major risk factor for development of cervical cancer. HPV 16 and HPV 18 have been considered as high-risk cancer associated HPV types. HPV types 31, 33, and 35 have been shown to have an intermediate relationship with cancer. These five HPV types together accounts for about 80% of cervical cancers. Additional high- and intermediate-risk HPV DNA types, including types 39, 45, 51, 52, 56, 58, 59 and 68, have been identified as the principal HPVs detectable in the remaining cancers. Low-risk HPV types 6 and 11 have been associated with the presence of genital warts. There are many other low-risk HPV types that are not associated with genital warts or cervical cancer. Until now, HPV cannot be cultured in vitro, and immunological tests are inadequate to determine the presence of HPV cervical infection. On the other hand, biopsies can be analyzed by nucleic acid hybridization to directly detect the presence of HPV DNA. Product Description Norgen’s HPV Low Risk (6/11) TaqMan PCR Kit is designed for the detection of HPV Low Risk (6/11) specific DNA in a real-time PCR based on the use of TaqMan technology. This kit is designed for research use only and not for use in diagnostic procedures. The detection of HPV Low Risk (6/11) specific DNA is based on TaqMan PCR providing a simple, reliable and rapid result for the detection of HPV Low Risk (6/11) infection. Norgen’s HPV Low Risk (6/11) TaqMan PCR Kit includes a PCR control to monitor for PCR inhibition, and to validate the quality of the sample and the detection result. The HPV Low Risk (6/11) TaqMan PCR Kit comprises Master Mix for the target and PCR control detection, Primer & Probe Mix, as well as a positive control and a negative control (nuclease-free water) to confirm the integrity of the kit reagents. The HPV Low Risk (6/11) TaqMan PCR Kit distinguishes between the detection of HPV 6 and HPV 11, allowing for clear and accurate results. HPV 11 detection is shown by amplification in the FAM channel. HPV 6 detection is shown by amplification in the Cy5 channel. Norgen’s HPV Low Risk (6/11) TaqMan PCR Kit was developed and validated to be used with the following PCR instruments: • Qiagen Rotor-Gene Q • BioRad CFX96 Touch™ Real-Time PCR Detection System • QuantStudioTM 7 Pro Real-Time PCR System Kit Components Component MDx TaqMan 2X PCR Master Mix HPV Low Risk (6/11) Primer & Probe Mix HPV Low Risk (6/11) Positive Control Nuclease-Free Water (Negative control) Product Insert

Storage Conditions and Product Stability • All kit components should be stored at -20°C upon arrival. • Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots. • All kit components can be stored for 2 years after the date of production without showing any reduction in performance. Customer-Supplied Reagents and Equipment Appropriate Real-Time PCR Instrument with FAM, HEX and Cy5 filter channel DNA Purification Kit o...

Do not substitute or mix reagents from different kit lots or from other manufacturers. Do not use components of the kit that have been stored for more than 2 years. The presence of PCR inhibitors may cause false negative or invalid results. Potential mutations within the target regions of the HPV Low Risk (6/11) genome covered by the primers in this kit may result in failure to detect the presence of the pathogen. Good laboratory practice is essential for the proper performance of this kit. Ensure that the purity of the kit and reactions is maintained at all times, and closely monitor all...

1. For each TaqMan PCR set, prepare one no template control PCR as shown in Table 1 below: Table 1. TaqMan PCR Negative Control Preparation PCR Components Target detection (with MDx TaqMan 2x PCR Master Mix) Nuclease-Free Water HPV Low Risk (6/11) Primer & Probe Mix Total Volume 2. Prepare the PCR reaction for sample detection as shown in Table 2 below. Table 2. TaqMan PCR HPV Low Risk (6/11) Assay Preparation PCR Components Target detection (with MDx TaqMan 2x PCR Master Mix) Nuclease-Free Water HPV Low Risk (6/11) Primer & Probe Mix Total Volume * The recommended amount of sample DNA to...

Table 4. HPV Low Risk (6/11) TaqMan PCR Program PCR Cycle D. HPV Low Risk (6/11) TaqMan PCR Assay Interpretation Table 5. Interpretation of Assay Results FAM (Target: HPV 11) For results obtained that are not covered in Table 5, please refer to the Frequently Asked Questions. Figure 1. Example of TaqMan PCR Positive result. Both PCR signals (FAM and Cy5) above the baseline from FAM, Cy5 and HEX channel indicate the successful PCR.

Figure 2. Example of TaqMan PCR Positive (Cy5) result. Both PCR signals above the baseline from Cy5 and HEX channel indicate the successful PCR. Figure 3. Example of TaqMan PCR Positive (FAM) result. Both PCR signals above the baseline from FAM and HEX channel indicate the successful PCR. Figure 4. Example of TaqMan PCR Negative result. No target DNA was detected in FAM and Cy5 channel but amplification signal from HEX indicates the successful PCR.