[1061] Simultaneous Detection, Typing and Quantification of All 15 High Risk HPV Types in Adeno-Squamous Carcinoma of the Uterine Cervix 
A Real Time PCR Based Study
MR Quddus, P Manna, CJ Sung, S Kerley, MM Steinhoff, WD Lawrence. Women & Infants Hospital, Brown University, Providence, RI; Physicians Reference Lab, Overland Park, KS
Background: Cervical adenosquamous carcinomas (ASqC) are a subtype of mixed glandular and squamous tumors with a genital HPV etiology. HPV 16 and 18 are regarded as the most common viral genotypes in cervical ASqC and have been reported in the following % proportions (HPV 16:18): 25:75; 33:33; 50:50 75:25. Multiple HPV types have been noted in pure cervical adenocarcinoma, but not in cervical ASqC. Type-specific detection of high risk HPV allows the detection of co-infection by multiple HPV serotypes as well as assessment of viral load per cell. We investigated the frequency of specific HPV serotypes in this largest series to date of cervical ASqC, and whether specific HPV serotypes and/or viral load per cell were associated with poor prognosis.
Design: 28 ASqC of the cervix were retrieved and a complex multiplex real time PCR test that simultaneously detects, types, and quantifies all 15 high risk HPV types was run. The test also detects and quantifies Beta-globin gene as internal control and also used to determine the normalized viral load (viral load/cell). A standard curve with the amplified products for each of the 15 high risk virus templates was plotted against the known standard concentration range of 10^0 to 10^7 to determine viral load. Prognosis-related histologic parameters of stromal invasion, VSI, LN and distant metastasis were correlated with the specific HPV serotypes and the viral copies.
Results:
| Multiple HPV types | Triple HPV Types | HPV 73 | HPV 31 | HPV 33 | HPV Other Than 16/18 | HPV 16/18 |
| 9 (32%) | 1 (3.5%) | 3 (11%) | 2 (7%) | 2 (7%) | 4 (14%) | 24 (86%) |
Poor prognosis histologic parameters e.g., +VSI, stromal invasion >50%, +LN, & vaginal/lung metastases were associated with i) 11 of 13 (84.6%) infected with HPV 16 ii) 6 of 13 (46.2%) cases of HPV 18, and iii) 4 of 4 cases of HPV 31, 45, 52, and 73 despite a low viral load. Viral loads range widely from 4 to 463906 copies per cell.
Conclusions: 32% of ASqC contain two or more high risk HPV types. 14.3% of cases are associated with NON-16/18 HPV types. Infection with 3 HPV serotypes is seen in 3.6% cases. Tumors with HPV 31, 45, 52 and 73 all have low (<500) viral loads but poor histologic prognostic parameters. HPV NON-18 is more frequently associated with a poor prognosis as compared to HPV 18. Viral load do not appear to correlate with poor prognosis histologic parameters.
Category: Gynecologic
Tuesday, March 10, 2009 2:00 PM
Platform Session: Section C, Tuesday Afternoon