Kim Chan Joo, Jeong Jeongmi Kim, Park Misun, Park Tae Shin, Park Tae Chul, Namkoong Sung Eun, Park Jong Sup
Division of Gynecologic Oncology and Department of Obstetrics & Gynecology, Kangnam St. Mary's Hospital, The Catholic University of Korea, Seoul, Korea.
Gynecol Oncol. 2003 May;89(2):210-7. doi: 10.1016/s0090-8258(02)00069-0.
In this study we examined the use of a new-human papillomavirus (HPV) detection method, the HPV oligonucleotide microarray system (Biomedlab Co., Korea), which we compared with the well-established HPV DNA detection system (Hybrid Capture II; HC-II, Digene Co.). This new method prompted us to develop a new HPV genotyping technique, using the oligonucleotide microarray, to detect the generic and type-specific sequence of HPV types. In particular, we undertook the evaluation of the clinical efficacy of the HPV oligonucleotide microarray for detecting HPV in cervical neoplastic lesions.
One hundred forty patients were involved and classified into three groups according to their histopathologic diagnoses: Group I (nonspecific chronic cervicitis; n = 61), Group II (low-grade squamous intraepithelial lesion (SIL); koilocytosis, and mild dysplasia; n = 39), and Group III (high-grade SIL; moderate, severe dysplasia and in situ carcinoma; n = 40). Cytological diagnoses were based on the Bethesda System and cervical samples were analyzed by the two methods. The HPV oligonucleotide microarray detected 15 types of high-risk HPV (HPV-16/-18/-31/-33/-35/-39/-45/-51/-52/-56/-58/-59/-66/-68/-69) and 7 types of low-risk HPV (HPV-6/-11/-34/-40/-42/-43/-44).
In 105 of the 140 cervical samples (75%), HPV DNAs were examined using the HC-II method. HPV detection rates using the HPV microarray agreed with those of HC-II. One HC-II-positive, but HPV microarray-negative, case occurred in the low-grade SIL (Group II) and was later confirmed negative for HPV. The other HPV microarray-positive but HC-II-negative case was found to be HPV-18 by PCR. Low-risk types of HPV were detected in 3 of 39 low-grade SIL cases (Group II) using the HPV microarray. HPV-16 was the most frequent type (32.1%) in all specimens tested, and was significantly more frequent in low-grade or high-grade intraepithelial lesions (Groups II or III) than in normal controls (Group I) (P < 0.05). HPV-58 was the second most common type (17.5%) in Group III. The HPV microarray was found to have advantages in terms of identifying the HPV genotypes and cases of multiple HPV infection. Double HPV infections were detected in 12 cases and triple HPV infections in 7 cases. Two cases were positive for four types of HPV (HPV-16/18/33/35, HPV-16/18/58/68). The sensitivity of HPV testing (HC-II; 94.9%, HPV microarray; 93.7%) for identifying patients with squamous intraepithelial lesion was significantly better than the sensitivity of cytology (77.1%, P < 0.05). On using multiple logistic regression analysis to estimate the relative risk of SIL versus HPV type, HPV-16-positive cases were found to have a 7.5-fold risk of SIL (95% CI = 3.28-16.51; P < 0.01). HPV-33 and HPV-58 were found to be significantly related to high-grade SILs (P < 0.01).
Our results suggest that the HPV oligonucleotide microarray is highly comparable to HC-II for detecting HPV in cervical specimens. The HPV oligonucleotide microarray provides useful information on viral genotype and multiple HPV infections in HPV-related cervical lesions. Genetic information on HPV in cervical specimens might be a particular benefit of the new procedure in the management of cervical neoplastic lesions
在本研究中，我们检测了一种新型人乳头瘤病毒（HPV）检测方法——HPV寡核苷酸微阵列系统（韩国Biomedlab公司），并将其与成熟的HPV DNA检测系统（杂交捕获二代；HC-II，Digene公司）进行比较。这种新方法促使我们开发一种利用寡核苷酸微阵列检测HPV通用序列和型特异性序列的新型HPV基因分型技术。特别是，我们对HPV寡核苷酸微阵列检测宫颈肿瘤病变中HPV的临床疗效进行了评估。
纳入140例患者，根据组织病理学诊断分为三组：第一组（非特异性慢性宫颈炎；n = 61），第二组（低级别鳞状上皮内病变（SIL）；挖空细胞和轻度发育异常；n = 39），第三组（高级别SIL；中度、重度发育异常和原位癌；n = 40）。细胞学诊断基于贝塞斯达系统，宫颈样本采用两种方法进行分析。HPV寡核苷酸微阵列检测15种高危型HPV（HPV-16/-18/-31/-33/-35/-39/-45/-51/-52/-56/-58/-59/-66/-68/-69）和7种低危型HPV（HPV-6/-11/-34/-40/-42/-43/-44）。
140例宫颈样本中的105例（75%）采用HC-II方法检测HPV DNA。HPV微阵列的HPV检测率与HC-II的检测率一致。在低级别SIL（第二组）中有1例HC-II阳性但HPV微阵列阴性的病例，后来经确认HPV为阴性。另一例HPV微阵列阳性但HC-II阴性的病例经PCR检测为HPV-18。采用HPV微阵列在39例低级别SIL病例（第二组）中的3例检测到低危型HPV。HPV-16是所有检测样本中最常见的类型（32.1%），在低级别或高级别上皮内病变（第二组或第三组）中比正常对照（第一组）更常见（P < 0.05）。HPV-58是第三组中第二常见的类型（17.5%）。发现HPV微阵列在鉴定HPV基因型和多重HPV感染病例方面具有优势。检测到12例双重HPV感染和7例三重HPV感染。2例为四种HPV类型阳性（HPV-16/18/33/35，HPV-16/18/58/68）。HPV检测（HC-II；94.9%，HPV微阵列；93.7%）用于鉴定鳞状上皮内病变患者的敏感性显著高于细胞学检查的敏感性（77.1%，P < 0.05）。使用多元逻辑回归分析评估SIL与HPV类型的相对风险时，发现HPV-16阳性病例发生SIL的风险为7.5倍（95%CI = 3.28 - 16.51；P < 0.01）。发现HPV-33和HPV-58与高级别SIL显著相关（P < 0.01）。
我们的结果表明，HPV寡核苷酸微阵列在检测宫颈样本中的HPV方面与HC-II具有高度可比性。HPV寡核苷酸微阵列为HPV相关宫颈病变中的病毒基因型和多重HPV感染提供了有用信息。宫颈样本中HPV的基因信息可能是这种新方法在宫颈肿瘤病变管理中的一个特别优势。
