Hybrid Capture System HPV DNA Test as a Screening Tool of Cervical Cancer / 대한부인종양콜포스코피학회잡지

OBJECTIVE: Although the use of cytology in screening has reduced morbidity and mortality from invasive cervical cancer, there has been limitation because of the low sensitivity and high false negative rate in the detection of preclinical or clinical disease of the cervix. This study was performed to explore the usefulness of Hybrid Capture System HPV DNA test as a diagnostic method of the cervical cancer. METHODS: Cervical cytology and Hybrid Capture System 1 (HCS1) HPV DNA test were performed in 1,023 women who visited Gynecologic Oncology Clinic in Guro Hospital, Korea University Medical Center. HPV DNA were retested by HCS1, HCS2 and Polymerase Chain Reaction(PCR), in randomly selected 200 women to compare these DNA tests. RESULTS: While the sensitivity and specificity of cervical cytology in detection of cervical lesion were 71.2% and 89.5%, those of HCS1 HPV DNA test were 63.0% and 86.9% respectively. There was significant difference in detection of cervical precancerous lesions between HCS1 and HCS2 (P<0.05), but not between HCS2 and PCR (P=0.14). And the sensitivity and specificity were improved with combining cytology and HCS1 HPV DNA test(82.2%, 86.9%~). CONCLUSION: HCS HPV DNA test may be considered as a useful adjunctive test with cervical cytology in cervical cancer screening.

Human Papillomavirus Typing by Polymerase Chain Reaction-Reverse Dot Blot Hybridization and Comparison with Hybrid Capture SystemTM / 대한임상병리학회지

BACKGROUND: Infection with certain Human Papillomavirus (HPV) type is strongly associated with the development of dysplasia and cancer of the cervix uteri. About 70 HPV types have been identified and some 25 of these have been found in the genital tract. HPV typing has diagnostic and prognostic importance to discriminate between 'low', 'intermediate', 'high' risk types. A Polymerase Chain Reaction-Reverse Dot Blot Hybridization (PCR-RDBH) method was developed for typing of HPV with consensus biotinylated primer generated PCR products in a single test. We attempted to know the clinical usefulness of PCR-RDBH and also compared PCR-RDBH with Hybrid captureTM system (HCS) method in same specimens. METHODS: HPV typing was performed on cervical swab samples obtained from 100 women with abnormal cervical cytology: 37 with Atypical Squamous Cells of Undetermined Significance (ASCUS), 14 with Low Grade Squamous Intraepithelial Lesions (LGSIL), 44 with High Grade Squamous Intraepithelial Lesions (HGSIL) and 5 with invasive carcinoma of the uterine cervix. HPV PCR screening was tested with consensus biotinylated primer. If HPV PCR screening was positive, RDBH was done for the typing of HPV. In RDBH, biotinylated PCR product was used in hybridization with a membrane on which 12 different oligonucleotide probes (type 6/11/16/18/ 31/33/35/39/45/51/52/56) of genital HPV types had been immobilized. Hybrid captureTM system (HCS, Digene Diagnostics) was used for screening of HPV. RESULTS: Of 100 abnormal cervical cytology specimens, the positivity of HPV PCR screening was 67%. In 52 specimens, HPV could be typed by RDBH. Type 16 was the most frequent and mixed infection was found in 6 cases, all combined with type 16. Among the 13 cervical cancer specimens confirmed by biopsy, 12 specimens was found to be infected high and intermediate risk types of HPV. In cervical swab, there was signifincant discrepancy in positivity of HPV infection between PCR-RDBH and HCS method. In 51 cases, negative for PCR-RDBH, 16 cases (31%) were positive by HCS. In 49 cases, positive for PCR-RDBH, 15 cases (31%) were negative by HCS. CONCLUSIONS: PCR-RDBH method can do HPV typing fast and easy with non-radioactive biotinylated primer in cervical swab specimens. It is shown to be useful method for HPV typing and have a high clinical applicability. The results between PCR-RDBH and HCS methods show a significant discrepancy, so further investigation is needed.

Clinical Implication and Evaluation and Management of Women with Low-Grade Cytologic Abnormality ( ASCUS/LSIL ) / 대한산부인과학회잡지

The evaluation of women with low-grade cytologic abnormalities including atypical cells of undetermined significance(ASCUS) and low-grade squamous intraepithelial lesion (LSIL) is a topic of considerable controversy. Some women with low-grade cytologic abnormalities will have high-grade intraepithelial neoplasia(CIN II or III) or even invasive cancer. Now, it is almost universally accepted that the majority of invasive cervical cancer and high-grade intraepithelial neoplasia are associated with Human papillomavirus(HPV). Because of this, there has been increasing tendancy on using another triaging methods for evaluating women with a low grade abnormal Papanicolaou smear. Traditionally colposcopy was used and recently HPV DNA test and cervicography was widely used as a triaging methods. This study used the data gained from 151 evaluable women with ASCUS and LSIL in the Department of Obstetrics and Gynecology, Catholic University Medical College, Kangnam St. Mary's Hospital from March 1995 to April 1996. 11,401 women had done cytology during this period and result was like this; 10,501 patients had normal cytology(92.1%), 244 patients had ASCUS(2.14%), 35 had AGCUS (atypical glandular cells of undetermined significance: 0.31%), 191 had LSIL(1.68%), 170 had HSIL(1.49%), and 160 had cervical cancers(1.41%). Patients with ASCUS were showed that CIN I WAS 5.6%(4/72), CIN II 4.2%(3/72) and CIN III 15.3%(11/72). Patients with LSIL were showed that CIN I was 5.1%(4/79), CIN II 7.6(6/79), CIN III 15.2%(12/79) and invasive cancer 7.6%(6/79). We adopted three management algorithms for evaluating women with ASCUS and LSIL. Three methods were like this; Colpscopy only or HPV DNA testing using hybrid capture which were used with DNAs extracted from the cerval swabs and tested with two pooled probes(probes containing HPV types of low oncogenic risk; 6, 11, 42, 43 and 44 or HPV types of high oncogenic risk; 16, 18, 31, 33, 35, 45, 51, 52 and 56) for HPV detection or both. We adopted only positive result with high risk HPV-probes in this article. At ASCUS group, sensitivity and specificity were 90.0% ad 32.5% in 62 women with colposcopy and 71.4%, 75% in 28 women with Hybrid Capture System. 27 women with colposcopy and Hybrid Capture System showed that sensitivity was 100%, and specificity was 26.3%. At LSIL group, sensitivity and specificity were 92.6% and 22.7% in 71 women with colposcopy and 69.2%, 65.4% in 39 women with Hybrid Capture System, 37 women with colposcopy and Hybrid Capture System showed that sensitivity was 91.7%, and specificity was 12%. Our result indicate that patients with smears showing some of ASCUS and LSIL are evaluated as CIN I, II, III and even invasive cancer. Therefore as a triage methods, the combination with colposcopy and HPV DNA test in women referred for low-grade abnormal cytology would be used as a sensitive methods for the detection of CINs. However, low -specificity of this combination approach was also considered as problem. This approach provides another information to differentiating ASCUS and LSIL into high- or low-risk group of progression.

The Study of Low-risk and High-risk Human Papillomaviruses Infection in cervical Carcinomas and Cervical Instaepithelial Lesions with Hybrid Capture system / 대한산부인과학회잡지

The assocication between human papillomavirus infection (HPV) and cervical lesionhas been well established. for detection and typing human papilloma virus deoxyribonucleic acid in cervical tissues, Southern blot hybridzation and polymerase chain reaction are commonly regarded as reference standard methods. However it has the limitation includeing technical difficulty, safety, subjectivey in result interpretation. Recently the chemilnuminescent molecular hybridization assay method has been windely used and it has been known that it can detect less hybridization assay method has been windely used and it has been known that it can detect less then 1 pg of DNA in a 100n1 aliquot of a crude specimen. This study was perfomed to determine the usefulness of hybrid capture HPV DNA assay for detecting low-risk and high-risk human papillomaviruses in histologically confirmed normal, cervical intraepithelial lesion(CIN) and invasive squamous cell carcinoma of the cervix, and to compare the correlation among cervical cytology, hiopsy finding and HPVs infelction ,and to detirmine whether the additon of the hybrid capture test to cytologic studies would improve the ability to identify signifcant lesions. Cervical cytologic smears, hybrid capture HPVs DNA assay, and pucnh bhiopsies were performed on 78 women who have normal cervix(28cases),cervical intraepithelila lesion (24cases), and invasive squamous cell carcinoma(26cases). At first the probes for low-risk HPV(6,11,42,43,44) and the proves for high-risk HPV(16,18,31,33,35,45,51,52,56)were used and secondly retyping was done for HPV 16 abd 18 in high-risk HPV positive cases.the results obtained were as follows; 1. Low-risk HPVs infections were 14.3% and 8.3% in normal cervix and cervical intraepithelial lesion respectively. High-risk HPVs infection were 7.1%, 70.8% and 73.1% in normal cervix, cervical intraeithelial lesion and squamous cell carcinoma respecitively. These was highly significant corelation between positive high-risk HPVs test, cervical intraepithelial lesion and squamous cell carcinoma. 2. Positivities of low and high-risk human papillomaviruses in patients with negative cytologic result were 9.1% and 15.1% respectinvely. 3. In patients with high-risk human papillomaviruses infection, human papilloma viurs 16 and 18 types were detected in 47.4% and 13.2%, respectinvely, and both HPV 16 and 18 positive and negative detention were 15.7% 23.7% respectinvely. Among 12patinent of cervical intraepithelial neoplasia with high risk HPVs infection, HPV 16 was detected in 66.6% and HPV 18 in 16.7% Among 24 squamous cell carcinomas with high risk HPVs infection, HPV 16was detected in 41.7% and HPV 16 in 12.5% and both HPV and 18 in 25% 4. In patients without koilocytosis by cervical tytology, low and high-risk human papillomaviruses or both were positive in 30.7% 5. In patients with positive high-risk HPVs, sensitivitives of koilocytosis were 64% and 69% in histologic and cytologc diagnosis, and specificities and specifictites were 58.5% and 62% respectively. Above result suggest that detection for high-risk human papilloma viruses type by hybrid capture assay improves the management of cervical intraepithelial neoplasia and is more useful method over cervical cytology only.

The Study of Low-risk and High-risk Human Papillomaviruses Infection in cervical Carcinomas and Cervical Instaepithelial Lesions with Hybrid Capture system / 대한산부인과학회잡지

The assocication between human papillomavirus infection (HPV) and cervical lesionhas been well established. for detection and typing human papilloma virus deoxyribonucleic acid in cervical tissues, Southern blot hybridzation and polymerase chain reaction are commonly regarded as reference standard methods. However it has the limitation includeing technical difficulty, safety, subjectivey in result interpretation. Recently the chemilnuminescent molecular hybridization assay method has been windely used and it has been known that it can detect less hybridization assay method has been windely used and it has been known that it can detect less then 1 pg of DNA in a 100n1 aliquot of a crude specimen. This study was perfomed to determine the usefulness of hybrid capture HPV DNA assay for detecting low-risk and high-risk human papillomaviruses in histologically confirmed normal, cervical intraepithelial lesion(CIN) and invasive squamous cell carcinoma of the cervix, and to compare the correlation among cervical cytology, hiopsy finding and HPVs infelction ,and to detirmine whether the additon of the hybrid capture test to cytologic studies would improve the ability to identify signifcant lesions. Cervical cytologic smears, hybrid capture HPVs DNA assay, and pucnh bhiopsies were performed on 78 women who have normal cervix(28cases),cervical intraepithelila lesion (24cases), and invasive squamous cell carcinoma(26cases). At first the probes for low-risk HPV(6,11,42,43,44) and the proves for high-risk HPV(16,18,31,33,35,45,51,52,56)were used and secondly retyping was done for HPV 16 abd 18 in high-risk HPV positive cases.the results obtained were as follows; 1. Low-risk HPVs infections were 14.3% and 8.3% in normal cervix and cervical intraepithelial lesion respectively. High-risk HPVs infection were 7.1%, 70.8% and 73.1% in normal cervix, cervical intraeithelial lesion and squamous cell carcinoma respecitively. These was highly significant corelation between positive high-risk HPVs test, cervical intraepithelial lesion and squamous cell carcinoma. 2. Positivities of low and high-risk human papillomaviruses in patients with negative cytologic result were 9.1% and 15.1% respectinvely. 3. In patients with high-risk human papillomaviruses infection, human papilloma viurs 16 and 18 types were detected in 47.4% and 13.2%, respectinvely, and both HPV 16 and 18 positive and negative detention were 15.7% 23.7% respectinvely. Among 12patinent of cervical intraepithelial neoplasia with high risk HPVs infection, HPV 16 was detected in 66.6% and HPV 18 in 16.7% Among 24 squamous cell carcinomas with high risk HPVs infection, HPV 16was detected in 41.7% and HPV 16 in 12.5% and both HPV and 18 in 25% 4. In patients without koilocytosis by cervical tytology, low and high-risk human papillomaviruses or both were positive in 30.7% 5. In patients with positive high-risk HPVs, sensitivitives of koilocytosis were 64% and 69% in histologic and cytologc diagnosis, and specificities and specifictites were 58.5% and 62% respectively. Above result suggest that detection for high-risk human papilloma viruses type by hybrid capture assay improves the management of cervical intraepithelial neoplasia and is more useful method over cervical cytology only.

Clinical Efficacy of Human Papilloma Virus DNA Test Using Hybrid Capture System in Health Screening of Normal Women / 대한부인종양콜포스코피학회잡지

Cervical cancer and its precursors are caused principally, if not exclusively, by HPV infection and HPV DNA is found in more than 90% of cervical cancers. Cervical cytology is limited by its false negativity and this may be supplimented by other adjunctive test such as HPV test. It is therefore important to explore the use of HPV DNA detection as a primary or supplementary screening method and to determine whether HPV typing can be used as a predictor of a lesion's clinical behavior. Cervical cytology and Hybrid Capture test for HPV detection were performed in 450 asymtomatic wornen visited Health Care Center in Kangnam St. Mary's Hospital,and none of whom was believed to have current cervical disease. The Papanicolaou cytology results were classified by The Bethesda System : 333(74.%) women were classified to within normal limit, 19(4.2%) benign reactive change, 38(8.4%) ASCUS, 59(13%) low grade SIL, and only one woman high grade SIL. Twenty five of 450(5.6%) women showed HPV infection by Hybid Capture test. Among 98 wornen with abnormal Papanicolaou cytology, 16(16.3%) women showed HPV DNA positivity. (continue)