ANALYSIS OF CYTOMORPHOLOGICALLY ABNORMAL CERVICAL SCRAPES FOR THE PRESENCE OF 27 MUCOSOTROPIC HUMAN PAPILLOMAVIRUS GENOTYPES, USING POLYMERASE CHAIN-REACTION

The aim of this study was to investigate the distribution of 27 mucoso tropic human papillomavirus (HPV) genotypes (HPV 6, 11, 13, 16, 18, 30 , 31, 32, 33, 35, 39, 40, 42, 43, 44, 45, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61 and 66) in cytomorphologically abnormal cervical scrapes (P ap IIIa-Pap IV; n = 1,373) using the polymerase chain reaction (PCR) m ethod on crude cell suspensions. The scrapes were analyzed for the pre sence of HPV DNA by HPV general-primer-mediated PCR (GP-PCR), which al lows the detection of a broad spectrum of HPV types at the subpicogram level. Subsequently, 2 HPV typing procedures based on either type-spe cific PCR (for HPV 6, 11, 16, 18, 31 and 33) or characterization of GP -PCR products by hybridization (for HPV 13, 30, 32, 35, 39, 40, 42, 43 , 44, 45, 51, 52, 53, 54, 55, 56, 57, 58, 59, 61 and 66) were applied. Increasing total HPV prevalence was found with increasing severity of dysplasia from 71% in Pap IIIa to 100% in Pap IV scrapes (carcinoma i n situ). The scrapes which were positive by type-specific PCR included 47% cases of Pap IIIa, 71% cases of Pap IIIb and 90% cases of Pap IV. Moreover, 12% of Pap IIIa scrapes, 6% of Pap IIIb scrapes and 8% of P ap IV scrapes revealed positivity for one or more of the remaining HPV types, as determined by successive hybridizations of the GP-PCR produ cts. Taking the typing data together, we noted that the level of HPV h eterogeneity decreased from 22 different HPV types (HPV 6, 11, 16, 18, 31, 33, 35, 39, 40, 42, 43, 44, 45, 51, 52, 54, 55, 56, 58, 59, 61 an d 66) detected in the group of Pap IIIa scrapes to 13 (HPV 6, 11, 16, 18, 31, 33, 35, 45, 51, 52, 58, 59 and 61) and 10 HPV genotypes (HPV 6 , 16, 18, 31, 33, 45, 51, 52, 54 and 58) in the Pap IIIb and Pap IV cl asses, respectively. An increasing prevalence rate from Pap IIIa to Pa p IV was found for HPV 16, 18, 31, 33, 45 and 54. The prevalence rate of identified HPV genotypes increased from 59% in Pap IIIa to 98% in P ap IV, indicating that almost all high-risk HPV genotypes related to c ervical cancer in The Netherlands have been characterized. (C) 1994 Wi ley-Liss, Inc.