자궁경부의 인유두종 바이러스 검출에 있어 HPV DNA chip 검사의 임상적 유용성

이제훈 ( Je Hoon Lee ),이기헌 ( Ki Heon Lee ),이인호 ( In Ho Lee ),우혁준 ( Hyuk Jun Woo ),성석주 ( Seok Ju Seong ),김태진 ( Tae Jin Kim ),임경택 ( Kyung Taek Lim ),심재욱 ( Jae Uk Shim ),박종택 ( Chong Taik Park ) 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.10

목적: 비정상 자궁경부 세포검사 소견을 보인 환자 중 질확대경 검사나 원추절제술을 통해 조직학적 병변의 유무가 확인된 환자들을 대상으로 HC-II와 HPV DNA chip 검사를 시행하여 각 검사의 임상적 유용성을 확인하고자 한다. 연구 방법: 2004년 7월부터 10월까지 비정상 자궁경부 세포검사 결과로 본원 부인종양센터에서 질확대경하 조직검사 또는 원추절제술을 시행하여 조직학적 소견이 확인된 200명의 환자에서 HPV 감염을 확인하기 위해 HC-II와 HPV DNA chip 검사를 동시에 시행하여 HPV 검출에 두 검사방법의 일치율을 비교하고, HPV DNA chip의 결과로 확인된 HPV 아형들을 분석하였다. 결과: 200명의 대상 환자 중 HC-II와 HPV DNA chip의 결과가 일치한 경우는 171명 (85.5%) 이었다. 비정상 자궁경부세포검사를 보인 환자 중 조직 검사 상 편평콘딜로마 이상의 병변이 확인된 111명의 환자에서, HC-II와 HPV DNA chip 검사의 민감도는 각각 91.0%와 88.3%였다. HPV DNA chip 검사에서 검출된 HPV의 아형들은 HPV-16이 가장 흔하게 검출되었고 (14.7%), 그 다음으로 HPV-58과 HPV-18이 14.1%와 9.8%의 순으로 검출되었다. 결론: 자궁경부의 병변에서 HPV의 검출에 사용되는 HPV DAN chip 검사는 HC-II에 견줄만한 민감도를 갖는 효과적인 방법이다. 또한 HPV DNA chip은 감염된 HPV의 아형을 확인하고 중복감염 등 임상적으로 유용한 정보를 제공하는 장점을 가지고 있다. Objective: This study was performed to investigate the efficacy of HPV DNA chip method for detection and genotyping of various human papillomavirus in the patients with intraepithelial lesions of uterine cervix. Methods: The study subjects included two hundred patients with abnormal Pap smear from July 2004 to October 2004. After confirmed the pathological status of the cervix with colposcopic biopsy or conization, we evaluated for HPV infection and genotyping with the commercially available Hybrid-Capture II assay (HC-II) and HPV DNA chip. Then we compared the concordance rate between the two methods for the detection of HPV and analysed the HPV genotypes. Results: We compared the results in HPV DNA chip with those in HC-II. In result, the concordance rate between the two methods for the detection of HPV was 85.5% (171 of 200 cases). In 111 patients confirmed the presence of lesions higher than flat condyloma in cervix by pathologc examination, sensitivities of HC-II and HPV DNA chip in detecting HPV were 91.0% and 88.3%, respectively. In HPV DNA chip, HPV-16 was the most frequent type (14.7%) in all patients, the next frequent types were HPV-58 (14.1%) and HPV-18 (9.2%). Conclusion: We confirmed that HPV DNA chip method was as sensitive and effective method for detecting HPV in cervical lesions as HC-II. And that it would provide useful clinical information on genotyping and multiple infections of HPV.

비성기부 지루각화증에서 HPV DNA Chip을 이용한 Human Papillomavirus DNA 검색

이운하 ( Un Ha Lee ),최우석 ( Woo Seok Choi ),김성우 ( Sung Woo Kim ),박현수 ( Hyun Su Park ),장상재 ( Sang Jai Jang ),황선욱 ( Seon Wook Hwang ),이혜경 ( Hye Kyung Lee ) 대한피부과학회 2008 大韓皮膚科學會誌 Vol.46 No.10

Background: The precise etiology of seborrheic keratosis (SK) is unknown. Genetics, sun exposure and infection have all been implicated as possible factors. Because of its clinical and histopathological similarities to verrucae vulgaris and condyloma acuminatum, human papillomavirus (HPV) has been suggested as a possible causative agent. In the previous studies, HPV were frequently detected in the genital lesions or hair follicles of immunocompromised hosts. Objective: A newly introduced HPV detection technique, the HPV DNA Chip analysis, contains 24 HPV probes and it has the advantage of being able to detect 24 types of HPV at once. The purpose of this study was to evaluate the presence of HPV DNA in the nongenital SK of immunocompetant individuals. Methods: We analyzed 31 biopsy specimens that were taken from patients with nongenital SK, and these specimens were compared with genital warts (the positive control) and distilled water in place of DNA (the negative control) with using HPV DNA Chip analysis and a polymerase chain reaction-based DNA microarray system as the HPV genotyping method. Results: By polymerase chain reaction (PCR), HPV DNA was detected in 2 of the 31 nongenital SK biopsies (6.5%). HPV DNA Chip analysis revealed that 3 of 31 nongenital SKs (9.7%) contained HPV DNA. Two distinct HPV genotypes were detected: HPV type 16 (n=2) and HPV type 42 (n=1). The duration of SK in the HPV positive group was longer than that of the SK in the negative group. The mean age of the patients in the HPV positive group was also older than the mean age of the negative group. There were no different histopathologic findings between the HPV positive and negative SK. Conclusion: This study did not provide any concrete evidence that HPV infection might directly play a part in the pathogenesis of nongenital SK. However, two distinct HPV DNA types were identified as types that have never been reported before. Further studies with a larger number of cases of SK are needed to confirm the presence of HPV DNA in nongenital SK and also to determine the role of HPV in the origin of nongenital SK. (Korean J Dermatol 2008;46(10):1321∼1327)

자궁경부의 인유두종바이러스 아형의 분석과 HPV DNA chip 검사의 임상적 유용성

최순호 ( Soon Ho Choi ),백경돈 ( Kyung Don Baik ),이수일 ( Su Il Lee ) 대한산부인과학회 2007 Obstetrics & Gynecology Science Vol.50 No.3

목적: 인유두종 바이러스 (human papillomavirus, HPV)는 현재 100여종의 아형이 발견되었지만 그 분포는 국가간 또는 지역간에 차이를 보이고 있다. 본 연구는 거주지가 부산 및 근교 지역인 여성의 자궁경부에 감염되어 있는 HPV 아형의 종류와 조직학적 검사와의 연관성을 분석하였다. 아울러 HPV DNA chip 검사가 세포검사의 단점을 보완하는 수단으로서 자궁경부 병변의 평가에 있어서 임상적 유용성이 있는지 알아보고자 하였다. 연구방법: 최근 3년간 자궁경부암 검사에서 비정상적 세포검사 소견을 나타내었던 환자 중, HPV DNA chip (바이오메드랩)을 이용한 검사에서 양성을 나타낸 143명을 대상으로 HPV 아형을 분석하였다. 조직검사를 하지 못한 28명을 제외한 115명을 대상으로 조직병리 소견과 HPV DNA검사의 결과를 함께 비교 분석하였다. 그리고 세포검사에서 비정형 세포및 저등급 병변을 나타낸 54명의 조직소견을 확인하고 고위험군 HPV의 검출을 조사하여 분석하였다. 결과: HPV 아형의 분포는 HPV 고위험군에서는 HPV 16형 42예 (26.1%), 58형 20예 (12.4%), 52형 16예 (9.9%), 35형 10예 (6.2%), 56형 9예 (5.6%), 51형 7예 (4.3%), 18형 6예 (3.7%) 등의 순이었고 HPV 저위험군에서는 HPV 6형 3예 (1.9%), 42형 3예 (1.9%), 11형 2예 (1.2%), 34형 2예 (1.2%) 등의 순이었다. 115명의 조직병리소견과 HPV DNA chip 검사의 결과와의 비교분석에서는 저위험군 HPV의 감염군은 모두가 저등급 병변을 보였으며 고위험군 HPV 감염군에서는 저등급 병변도 나타났지만 주로 고등급 병변을 보였다. 그리고 세포검사에서 비정형 세포및 저등급 병변의 소견을 보였던 54명 중 조직검사에서는 13명이 고등급 병변 이상을 나타내어서 세포검사의 고등급 병변에 대한 위음성률은 24.1%이었고 여기에는 침윤암도 2예 포함되었다. HPV 아형은 HPV 16형, 58형, 52형, 35형, 56형, 51형, 18형의 빈도순으로 검출되었다. 결론: 앞으로 여러 지역에서 더 많은 예를 대상으로 한 연구가 이뤄져 통합이 되어야 할 것으로 생각된다. 그리고 HPV DNA chip 검사는 보조적으로 사용하면 세포검사의 고등급 병변에 대한 위음성률을 낮추고 양성 예측도를 높여 자궁경부암으로 진행이 되기 전에 진단이 가능할 것으로 판단되어 임상적 유용성이 있다고 생각된다. Objective: Human papillomavirus (HPV) has been identified more than 100 HPV subtypes. The distributions of subtypes are different according to nations and regions. We analysed subtype of infection with HPV among women who live in Pusan and surburbs of Pusan. We accessed the clinical usefulness of HPV DNA chip test as a supplementary method of Pap smear in the evaluation of cervical lesion. Method: This study was undertaken from January 2002 to January 2005 and the samples were collected from the patients who had abnormal Pap smear. We analysed subtypes of 143 positive cases with HPV DNA chip (Biomedlab) test and estimated pathologic reports of 115 patients except 28 patients who had not biopsy. We investigated pathologic results of 54 of 115 patients who had atypical squamous cells / low grade squamous intraepithelial lesion (ASC/LSIL) in Pap smear and examined high risk HPV in 54 pathologic results. Results: The prevalence of HPV subtypes was 42 cases of HPV-16, 20 cases of HPV- 58, 16 cases of HPV-52, 10 cases of HPV-35, 9 cases of HPV-56, 7 cases of HPV-51, 6 cases of HPV-18 in descending order of incidence in high risk HPV group and 3 cases of HPV-6, 3 cases of HPV-42, 2 cases of HPV-34, 2 cases of HPV-43 in descending order of incidence in low risk HPV group. The results of HPV DNA chip test and 115 pathologic reports were estimated by comparative study. A pure infection with low risk HPV group was detected in low grade lesion. Infection with high risk HPV group was also detected in low grade lesion but was mainly detected in high grade lesion. The pathologic results of 54 patients who had ASC / LSIL in Pap smear were 13 patients had above high grade lesion include 2 cases of invasive carcinoma so false negative rate of Pap smear in the detection of high grade lesion was 24%. Conclusion: HPV subtypes were detected HPV 16, 58, 52, 35, 56, 51, 18 types in descending order of incidence and prevalence. Mass study and integrated data from larger population and various regions in many hospitals will be needed. And the supplementary use of HPV DNA chip test may provide clinical usefulness because it can reduce the false negative rate of Pap smear and improve the positive predictive value in the detection of high grade cervical lesion and it enables to decrease the incidence of cervical cancer.

Evaluation of human papillomavirus (HPV) genotyping assays using type-specific HPV L1 reference DNA

한경호 한국유전학회 2021 Genes & Genomics Vol.43 No.7

Background Human papillomaviruses (HPV) are known to play a central etiological role in the development of cervical cancer. General HPV genotyping methods consist of PCR with consensus primers combined with various detection methods. Objective The aim was to develop HPV L1 DNA reference materials to evaluate the sensitivity, specifcity, and accuracy of genotyping results obtained from the HPV DNA Genotyping Chip (HPV CHIP) and RFMP assays. Methods In this study, the Ministry of Food and Drug Safety (MFDS) established reference DNA materials for the L1 gene from 41 subtypes of anogenital HPV to aid in genotyping human papillomavirus (HPV) strains. Of these, 22 subtypes were obtained from cervical scrape samples of Korean women and 19 subtypes were synthesized. These reference materials include 13 high-risk types (HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68), 3 probable high-risk types (HPV-26, 53, and 66), 16 low-risk types (HPV-6, 10, 11, 27, 34, 40, 42, 43, 44, 54, 55, 61, 70, 72, 73, and 81), and 8 undetermined-risk types (HPV-3, 57, 62, 67, 69, 71, 74, and 84). After confrming the sequences by standard methods, these HPV L1 DNA reference materials were then used to compare results from the HPV DNA Genotyping Chip (HPV CHIP) and restriction fragment mass polymorphism (RFMP) assays. Results Data collected from the HPV CHIP and RFMP assay showed comparably high sensitivity and accuracy. Both assays could detect 102 or more copies/μl of HPV L1 DNA from 39 types of HPV, with higher accuracy in detecting samples with mixed types of HPV. Conclusion The present study confrms the HPV L1 DNA reference materials developed by MFDS are reliable and useful for the evaluation of HPV genotyping assays.

사람유두종바이러스의 검출과 유전자형 분석을 위한 HPV28 Detection 검사와 HPV DNA Chip 검사의 비교 평가

신은심,배효진,송완근,정선경,황유성 대한진단검사의학회 2013 Laboratory Medicine Online Vol.3 No.4

Background: The HPV28 Detection test (Seegene) is a real-time polymerase chain reaction assay that is designed for testing a total of 28 human papillomavirus (HPV) genotypes and estimating the approximate HPV viral load. The aim of this study was to evaluate the clinical applicability of the HPV28 Detection test with regard to the prevalence of HPV infection and distribution of HPV genotypes by using the HPV28 Detection and HPV DNA Chip tests (Biomedlab). Methods: HPV DNA Chip and HPV28 Detection tests were performed for 500 cervical swab specimens. HPV genotype results were confirmed by sequencing analysis of the specimens that showed discordant results in the 2 test methods. Results: The positive rate of HPV detection determined by using HPV28 Detection and HPV DNA Chip tests were 43.8% and 40.6%, respectively. The sequencing results in 64 discordant specimens that showed single HPV infection in the 2 test methods were in complete agreement with the test results obtained with the HPV28 Detection test. The genotyping results of the HPV28 Detection test were 100% concordant in repeated experiments with HPV-infected specimens that have 12 different HPV genotypes, i.e., types 16, 31, 33, 39, 42, 51, 52, 53, 58, 66, 68, and 70. The HPV28 Detection test was 100-fold more sensitive than the HPV DNA Chip test with serially diluted HPV DNAs. Conclusions: The HPV28 Detection test can be applied in the clinical field as an HPV genotyping test can accurately identify various HPV genotypes with high specificity and low detection limit.

질 확대경상 비정상 소견을 보인 환자에서 HPV DNA chip과 Hybrid-Capture II assay의 임상적 유용성 비교

김태정,정찬권,이아원,정은선,최영진,이교영,박종섭,Kim, Tae-Jung,Jung, Chan-Kwon,Lee, Ah-Won,Jung, Eun-Sun,Choi, Young-Jin,Lee, Kyo-Young,Park, Jong-Sup 대한세포병리학회 2008 대한세포병리학회지 Vol.19 No.2

This study was performed to compare the efficacy between a DNA chip method and a Hybrid-Capture II assay (HC-II) for detecting human papillomavirus in patients with intraepithelial lesions of the uterine cervix. From May, 2005, to June, 2006, 192 patients with abnormal colposcopic findings received cervical cytology, HC-II and HPV DNA chip tests, and colposcopic biopsy or conization. We compared the results of HC-II and HPV DNA chip in conjunction with liquid based cervical cytology (LBCC) and confirmed the results of biopsy or conization. The sensitivity of the HPV DNA chip test was higher than HC-II or LBCC. The HPV DNA chip in conjunction with LBCC showed higher sensitivity than any single method and higher sensitivity than HC-II with LBCC. We confirmed that the HPV DNA chip test was more sensitive for detecting HPV in cervical lesions than HC-II, and that it would provide more useful clinical information about HPV type and its multiple infections.

자궁 경부 인유두종 바이러스 감염에 대한 세포검사와 DNA 검사의 비교 연구

최경운,김활웅,김지연,이창훈,설미영 부산대학교 병원 암연구소 2006 부산대병원학술지 Vol.- No.20

Purpose: A newly introduced HPV detection technique in cervical intraepithelial lesion, the HPV DNA Chip test, contains 24 HPV probes and has the advantage of being able to detect 24 HPV types simultaneously. The authors performed HPV genotyping test in cervical specimens from September 2005 to March 2006, and compared their cytologic and histologic diagnoses. Subject and Methods: The HPV DNA Chip test was performed in samples of 263 patients. Among these, 49 patients underwent cervical smear cytology and biopsy. Results: In 160 cases (60.8%) of 263 cases, the HPV DNA was detected. HPV-16 (37.5%), HPV-58 (16.3%), HPV-33 (12.5%), and HPV-18 (10.0%) were frequently detected, including single and multiple infections. The multiple HPV infection was found in 31 cases (11.8%). The sensitivity (91.7%) of the HPV genotyping test using the DNA chip method for detecting HSIL or worse was superior compared with the sensitivity (79.2%) of cytologic diagnosis. Conclusions: Our results indicate that HPV DNA chip test can overcome the low sensitivity of cytologic test and improve the sensitivity of the screening test for cervical intraepithelial lesion.

Cytomorphologic Features According to HPV DNA Type in Histologically Proven Cases of the Uterine Cervix

최인호,이소영,이동화,김동원,진윤미 대한병리학회 2011 Journal of Pathology and Translational Medicine Vol.45 No.6

Background: This study investigated whether human papillomavirus (HPV) genotype is related to koilocytic changes in cervical cytology and histology, and what factors cause discrepancies among cytology, HPV DNA chip tests, and biopsies. Methods: We examined 174 of 949 cases histologically confirmed by both cytology and HPV DNA chip testing. We analyzed koilocytic changes in cytology and biopsies according to HPV genotype. Results: HPV-16 significantly coincided with nuclear size variation and hyperchromasia, although the cytomorphologic features correlated with other HPV genotypes were not statistically significant. By analyzing 68 cases in which there were discrepancies between the HPV DNA chip test and histological results, we confirmed that artifacts or glycogen acanthosis resulted in the over-diagnoses of four HPV-negative cases with normal cytology. Four diagnostic errors and four sampling errors were present in eight HPV-positive cases. The degree of nuclear size variation significantly influenced the cytologically under-diagnosed cases (p=0.006). Conclusions: Other than HPV-16, HPV genotype exhibited no cytological or histological differences. The discrepancy between the results of HPV DNA chip test and histology was created by glycogen acanthosis, immature squamous metaplasia, artifacts, and sampling errors.

Detection of Human Papillomavirus in Male and Female Urine by Electrochemical DNA Chip and PCR Sequencing

Nilyanimit, Pornjarim,Wanlapakorn, Nasamon,Niruthisard, Somchai,Pohthipornthawat, Natkrita,Karalak, Anant,Laowahutanont, Piyawat,Phanuphak, Nittaya,Gemma, Nobuhiro,Poovorawan, Yong Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.9

Background: Cervical cancer is the second most common cancer in Thai women after breast cancer. Currently, the Papanicolaou (Pap) smear is the recommended procedure for cervical cancer screening in Thailand, but only a relatively small percentage of women follow this screening program. An alternative method to detect HPV genotypes associated with cervical cancer is self-sampling of urine, which is a more widely accepted method. Our study aimed to evaluate the prevalence of HPV in Thai women using urine and cervical swabs and prevalence of HPV in Thai men using urine samples. Materials and Methods: Tumorigenic HPV detection was accomplished by electrochemical DNA chip and PCR/direct sequencing. In addition to HPV prevalence, we report the concordance between different methods and sample types. One-hundred and sixteen women and 100 men were recruited. Histological examination revealed normal cytology in 52 women, atypical squamous cells of undetermined significance (ASCUS) in 9, low-grade squamous intraepithelial lesions (LSIL) in 24, and high-grade squamous intraepithelial lesions (HSIL) in 31. One-hundred men were classified as heterosexuals (n=45) and homosexuals (n=55). Results: The most prevalent HPV genotype in our study was HPV16. The HPV detection rate was generally lower in urine samples compared with cervical samples. Overall, there was good agreement for the detection of carcinogenic HPV from female cervical samples between the DNA chip and PCR/sequencing, with 88.8% total agreement and a kappa value of 0.76. In male urine samples, the level of agreement was higher in heterosexuals compared with homosexuals. Conclusions: Further improvement is required to increase an overall yield of HPV DNA detection in urine samples before clinical application of a urine-based HPV screening program. The electrochemical DNA chip test is a promising technique for carcinogenic HPV detection.

자궁경부 종양 환자에 있어 인유두종바이러스 DNA chip 검사와 하이브리드캡쳐 Ⅱ 검사 간의 유용성 비교

권한성,김영태,김재욱,김성훈 대한부인종양 콜포스코피학회 2002 Journal of Gynecologic Oncology Vol.13 No.4

목적 : 현재까지 밝혀진 바에 따르면 인유두중 바이러스는 자궁경부암의 주요 인자로 알려져 있으며 저자들은 현재까지 알려진 검사법 중 하이브리도캡쳐 Ⅱ와 최근 각광받고 있는 oligonucleotide microarray법 간의 임상적 유용성을 알아보고 oligonucleotide microarray법을 통해 알 수 있는 인유두종바이러스의 다중 감염과 자궁경부 병변과의 상관 관계를 알아보고자 하였다. 연구 방법 : 2001년 12월부터 2002년 2월까지 본원에서 하이브리드캡쳐 Ⅱ, oligonucleotide microarray법, 세포진 검사를 시행받고, 이후 조직학적 진단을 받은 102명을 대상으로 각각의 검사법에 있어 자궁경부 병변에 대한 민감도, 특이도, 양성 예측도, 음성 예측도 등을 구하여 임상적 유용성을 비교하고자 하였다. 결과 : 저등급 상피내종양(LGCIN) 이상의 경우 하이브리드캡쳐 Ⅱ는 민감도 65.7%, 특이도 81.3%, 양성 예측도 85.5%, 음성 예측도 52.0%였고, oligonucleotide microarray법은 민감도 85.9%, 특이도 43.8%, 양성 예측도 77.2%, 음성 예측도 60.9%였다. 고등급 상피내종양(HGCIN) 이상의 경우 하이브리드캡쳐 Ⅱ는 민감도 71.7%, 특이도 71.4%, 양성 예측도 73.1%, 음성 예측도 70.0%였고, oligonucleotide microarray법은 민감도 86.8%, 특이도 34.7%, 양성 예측도 58.2%, 음성 예측도 73.9%였다. 두 검사법을 세포진 검사와 병행하였을 경우 특이도를 제외한 민감도, 양성 예측도, 음성 예측도가 향상된 소견을 보였다. 다중 감염일 경우 단독 감염일 때보다 자궁경부 병변의 빈도가 높았다. 결론 : 단독 검사로서 하이브리드캡쳐 Ⅱ에 비해 oligonucleotide microarray법이 더 높은 민감도를 보였고, 두 검사법을 세포진 검사와 병행하였을 때 screening performance가 향상된 소견을 보여 선별 검사로서 유용한 면이 있으며, 다중 감염일 때 저등급병변 이상의 발생 빈도가 높아 이에 대한 정밀한 추적 관찰이 요망된다고 하겠다. Objective : The detection of high-risk human papilloma virus (HPV) allows us to predict the presence and future development of cervical intraepithelial neoplasia. The aim of this study was to evaluate the efficacy and diagnostic performance of hybrid capture (HC) II and oligonucleotide microarray (WV DNA chip) in detecting high risk HPV. Methods : Cytologic study, HC II, HPV DNA chip test and pathologic study (punch biopsy, cone biopsy or hysterectomy) were performed on 102 patients. The screening performance was evaluated, such as sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Results : HPV tests were performed to detect high-risk types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68). The sensitivity of each test to detect low grade cervical intraepithelial neoplasia (LGCIN), high grade cervical intraepithelial neoplasia (HGCIN) and invasive cancer was 65.7% in HC II and 85.7% in oligonucleotide microarray. The combination of each test and cytologic study resulted in higher sensitivities, 90.0% in HC II and 95.5% in oligonucleotide. For patients with HGCIN and invasive cancer, the sensitivity was 71.7% in HC II and 86.8% in oligonucleotide microarray. In cases of multiple HPV subtype infection, the PPV of DNA chip was 89.5%. Conclusion : DNA chip test was more sensitive in detecting women with CIN lesion/invasive cancer and high risk HPV infection than HC II in a single test but no significant difference is found when combined with cytologic study. There was a highly significant correlation between multiple HPV subtype infection and CIN lesion/invasive cancer.