Journal List > Korean J Gynecol Oncol Colposc > v.1(1) > 1123599

Song and Ryu: Identification of Human Papillomavirus(HPV) in Patients with Cervical Cancer by DNA Hybridization

Abstract

Genomic DNAs were extracted from cervical lavages of 49 patients with cervical cancer. Dot and Southern blot hybridization were performed using the P-labeled HFV DNA probes to find high risk HPV(type 16 and 18) infection that is known as the mast prevalent pathogenic factor in cervical cancer. Furthermore, genornic DNAs purified frnm cervical cancer tissues were studied in 8 out of 49 patients allowing us to convince the results from cervical lavages. The results were as follaws: 1. Dot blot analysis were used to examine the sensitivity and specificity of hybridization condition and HPV-DNA probes. Fasitive signals were obtained even at the level of 10pg for HPV DNA, but no signals could he detected at the level of as much as 400pg for salmon sperm DNA. 2. Dot blot of DNAs from cervircal lavages showed positive signals in 32.7%(16/49) with HPV type 16 probe and 20.4% (10/49)and one mixed infection was found. 3. When the DNAs from cervircal lavages of 49 patients were classified according to the clinical stage of cervical cancer, the infection rates of HPV type 16 and 18 were 50% (2/4) in CIN, 80% (4/5) in stage I, 64. 2% (9/14) in stage I b, 45% (9I20) in stage II and 16. 7% (1/6) in stage Ill and K respectively. The occurrenr,e of HPV type 16 and 18 seemed to be the highest in the cervical cancer stage 1 (68.4%(13/19). 4. Experiments perfornecl with genomic DNAs from 8 cancer tissues showed similar results compared to those of cervical lavages, but the intensity of positive signals was stronger. 5. Genomic DNAs from 5 patients(3 cases from cervical lavages and 2 cases from cervical cancer tissues) which showed strong positive signals to the dot blot analysis were further examined by Southern blot hybridixation using HFV type 16 DNA probe. When DNAs were digested with Pst 1 restriction enzyme, the five characteristic frgmenta of BFV type 16(2.8, l.9, l.6, 1.0 and 0.5 kb long in length) were recognized in ell 5 cases, These results may suggest a direet relatianship between HPV type 16 & 18 infectioas considered as the most effective methods for HPV detectioe and typing. Mo1ecular biclogieal studies in the reserarch of HPV are expected to reveal and help us understand the pathogenesis of cervical cancer.

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