USE OF MULTIPLE PCR PRIMER SETS FOR OPTIMAL DETECTION OF HUMAN PAPILLOMAVIRUS

Using multiple PCR primer sets, we tried to optimize the detection of human papillomavirus (HPV) in DNA samples isolated from 361 frozen bio psy specimens from patients with invasive cervical carcinomas. The HPV s detected were placed into three distinct groups, including group I/I -neg at Telelab (Skien, Norway) and group I-neg and group II at the No rwegian Radium Hospital (Oslo, Norway). The consensus primer sets were Oli-1b-oli-2i, My09-My11, Gp5-Gp6, and Gp5+-Gp6+ from the HPV L1 gene and CpI-CpIIG from the E1 gene. Using these consensus primers togethe r with the type-specific primers from E6-E7, we found that 355 patient s (98%) were HPV positive. Type-specific primers for HPV types 11, 16, 18, 31, 33, and 35 detected more HPV-infected patients than the most sensitive consensus primer set, while the three consensus primer sets My, Gp/Gp+, and Cp together detected more HPV-positive patients than t he type-specific primers. Testing of sensitivity of the PCR with SiHa cells serially diluted in lymphocytes (HPV-negative cells) indicated a detection limit of 6,300 HPV type 16 DNA copies with consensus primer s (My, Gp+, and Cp) and 126 original HPV type 16 DNA copies with type- specific primers. Comparison of the amplification results for consensu s L1 primers and type-specific E6-E7 primers indicated the presence of L1 deletions in 23 of 56 samples. The conclusion is that in PCR detec tion systems, multiple consensus primers and type-specific primers sho uld be used in order to detect all patients harboring HPV.