Introduction
Over 130 HPV types have been documented in the literature, approximately 40 of which infect the anogenital area and are sexually transmitted. Persistent anogenital high-risk HPV infection is associated with the majority of cervical cancers. Cervical cancer is highly preventable when cytological and HPV screening programs are employed to facilitate early detection and treatment of pre-cancerous lesions. Of the sexually transmitted HPV genotypes, 14 (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) are considered high risk as they lead to cervical cancer or anogenital warts. In addition, HPV types 6 and 11 are considered low risk for cervical cancer. However, they are the cause of 90% of all cases of genital warts as well as respiratory papillomatosis.
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Principle of Assay
The crb® HPV Complete qPCR uses probe-based qPCR technology to detect HPV with high specificity. Each sequence-specific probe contains a 5’ fluorophore and a 3’ quencher. On its own, the probe’s 3’ quencher effectively absorbs the emission from the nearby 5’ fluorophore so that no net fluorescence is detected. During PCR amplification, the probe binds to its specific target template. While replicating the DNA sample, the PCR Taq Polymerase’s 5’ to 3’ exonuclease activity also hydrolyzes the probe, releasing a free-floating 5’ fluorophore that is detectable by the qPCR machine and can be translated into an amplification plot. This HPV Complete qPCR kit identifies 14 types of high-risk HPV, 2 types of low-risk HPV, and a human ACTIN control using 4 fluorescent channels in one tube: ROX, FAM, HEX, and Cy5. As a result, this kit can not only detect the 14 high-risk HPV types and 2 low-risk HPV types, it is also capable of distinguishing HPV 16 and 18 from all the other types. The internal control, human ACTIN, indicates the quality of extracted DNA.
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