Rapid Test for Identification of a Human Papillomavirus 16 E6 L83V Variant


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Abstract

A variant of human papillomavirus (HPV) 16 has been shown recently to be more prevalent in invasive cervical carcinoma than in preinvasive lesions. This HPV 16 variant possesses a common mutation (T to G) in nucleotide 350 (codon 83) of the E6 gene, resulting in an amino acid shift, L83V, in the E6 protein. This mutation was believed to signify preinvasive cervical lesions with a high probability of progression to invasive carcinoma. The purpose of the present investigation is to describe a rapid method for the detection of this variant HPV 16, E6 (L83V). Paraffin blocks of 18 gynecologic biopsy specimens were collected, all displaying the morphology of cervical intraepithelial neoplasia (CIN I-III) and a positive HPV 16 test. Sections from these blocks were used for DNA extraction. A DNA sequence of the E6 gene containing 176 bases (including codon 83) was amplified by polymerase chain reaction (PCR) and analyzed by non-radioactive single-strand conformational polymorphism (SSCP) analysis. A divergent SSCP pattern was observed in 7 of the HPV 16 positive biopsy specimens. A DNA sequence analysis of the PCR products revealed the conversion of Leu to Val in codon 83 of the E6 gene, correlating to the divergent band pattern. This PCR-SSCP method can be used to test for HPV 16 in women who are at serious risk of developing invasive cervical carcinoma.

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