| Detection of Cervical HPV Infection Using Reverse Dot Blot Hybridization Technique. |
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Y T Kim, S K Kim, J H Kim, J S Jung, J H Na, C W Ko, Y M Kim, C H Nam, J E Mok |
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| Abstract |
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The aims of the study were to determine the diagnostic performance of the new reverse dot blot hybridization (RDBH) technique in HPV detection and typing and to compare it with that of Hybrid Capture systemTM (HCS) in Korean women with preinvasive and invasive cervical disease. Cervical swabs from 100 women with abnormal cytology (37 with ASCUS, 14 with LSIL, 44 with HSIL and 5 invasive squamous cell carcinoma of the uterine cervix) were assayed for high-or intermediate-risk HPV DNA by both HCS and PCR followed by RDBH. HPV screening was done by PCR amplifying HPV L1 product with consensus primers followed by non-radioactive reverse dot blot hybridization as a single test. Biotin-14-dATP was incorporated into the amplified HPV DNA, which was then used as a single probe in hybridization with a membrane on which eight high-or intermediate-risk genital HPV types had been immobilized (type 16, 18, 31, 33, 35, 45, 51, 56). This results were as follows; The overall HPV DNA positivity was 49.0% (49/100) by RDBH. The HPV DNA positivities according to cytology were 24.3% (9/37) in ASCUS, 35.7% (5/14) in LSIL, 70.4% (31/44) in HSIL and 80% (4/5) in invasive cancer resulting significant correlation between grade of abnormal cytology and HPV DNA positivity. The HPV DNA positivities according to histology were 60.0% (3/5) in CIN I or II, 58.1% (25/43) in CIN III and 92.3% (12/13) in invasive cancer resulting significantly high HPV DNA positivity in invasive cancer. Type 16 was most frequent and mixed infection was found in 6 cases, all combined with type 16. There was a significant correlation between the positivity of HPV type 16 or 18 and the grade of cervical lesion (p<0.05). However, there is significant discrepancy in positivity of HPV infection between the HCS and RDBH methods. In 50 cases positive for high-or intermediate-risk HPV by the HCS, 15 cases (32.0%) were negative by RDBH. In another 50 cases negative for high-or intermediate-risk HPV by the HCS, 15 cases (30.0%) were positive by RDBH. Therefore, further investigation in large scale and confirmation by southern blotting or PCR typing are needed to explain the discrepancy between HCS and RDBH methods. In conclusion, reverse dot blot hybridization technique is fast and easy method to detect and type HPVs by a single test and may have high clinical applicability. |
| Key Words:
Human papillomavirus, Reverse dot blot hybridization, CIN, Cervical cancer |
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