Evaluation of a commercialized in situ hybridization assay for detecting human papillomavirus DNA in tissue specimens from patients with cervical Intraepithelial neoplasia and cervical carcinoma
Evaluation of a commercialized in situ hybridization assay for detecting human papillomavirus DNA in tissue specimens from patients with cervical Intraepithelial neoplasia and cervical carcinoma
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DOI：
10.1128/jcm.01299-07
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发表时间：
2008-01-01
影响因子：
9.4
通讯作者：
Sneigel, Nour
中科院分区：
文献类型：
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作者：
Guo, Ming;Gong, Yun;Sneigel, Nour
为了评价商业化的原位杂交（ISH）检测人乳头瘤病毒（HPV）DNA的能力，我们比较了一种新的ISH探针Inform HPV III（Ventana Medical Systems，Tucson，AZ）的方法，而不是PCR测定以检测具有正常子宫颈的子宫颈组织样本中的HPV DNA宫颈上皮内瘤变（CIN）20例，CIN 1型27例，CIN 2型28例，CIN 21型33例，宫颈癌29例。一般HPV DNA检测使用共识引物介导的PCR检测。使用EasyChip HPV印迹（King Car Yuan Shan Institute，I-Lan，Taiwan）进行HPV基因分型。采用实时荧光定量PCR检测HPV 16整合状态（E2/E6比值）。我们的研究结果表明，ISH和PCR在所有CIN类别中检测HPV DNA的一致性良好，无显著差异（Kappa系数，0.34至0.63; P=0.13至1.0）。然而，ISH检测到的HPV阳性病例明显少于PCR（Kappa系数，0.2; P=0.03）。11例ISH-PCR+结果的病例具有可通过Inform HPV III检测的HPV类型。5例ISH-PCR+的癌组织中HPV 16的整合水平显著高于ISH+的癌组织（P=0.008）。因此，游离型HPV 16在癌组织中的低拷贝数可能是ISH假阴性结果的原因。尽管HPV的点状信号模式随着疾病的严重程度而显著增加（P趋势=0.01），但在仅具有点状信号的病例与具有混合点状和弥漫性信号的病例之间没有观察到HPV 16整合状态的显著差异（P=0.4）。总之，使用Inform HPV III探针的ISH在检测CIN宫颈组织中的HPV DNA方面似乎与PCR相当。假阴性ISH结果似乎与附加型HPV 16的较低拷贝数相关，但与Inform HPV III探针检测特定HPV类型的能力无关。此外，信号模式，特别是混合点状和弥漫性HPV模式，不能可靠地用于预测病毒整合状态。
To evaluate a commercialized in situ hybridization (ISH) assay for detecting human papillomavirus (HPV) DNA, we compared the ability of a new ISH probe, Inform HPV III (Ventana Medical Systems, Tucson, AZ), to that of PCR assays to detect HPV DNA in cervical tissue specimens with normal cervix (20 cases), cervical intraepithelial neoplasia (CIN; CIN 1, 27 cases; CIN 2, 28 cases; and CIN 21, 33 cases), and cervical carcinoma (29 cases). General HPV DNA was detected using consensus primer-mediated PCR assays. HPV genotyping was performed by using EasyChip HPV blot (King Car Yuan Shan Institute, I-Lan, Taiwan). HPV16 integration status (E2/E6 ratio) was determined by using quantitative real-time PCR. Our findings showed that the ISH and PCR had fair to good agreements in detecting HPV DNA across all CIN categories without significant differences (Kappa coefficient, 0.34 to 0.63; P=0.13 to 1.0). However, ISH detected significantly fewer HPV-positive cases in carcinoma than PCR did (Kappa coefficient, 0.2; P=0.03). Eleven cases with ISH-PCR+ results had HPV types that can be detected by Inform HPV III. Five carcinoma cases with ISH-PCR+ results showed a significantly higher level of integrated HPV16 (P=0.008) than did the ISH+ cases. As a consequence, lower copy numbers of episomal HPV16 in carcinoma might be the cause for the false-negative ISH results. Although the punctate signal pattern of HPV significantly increased with the severity of disease (P trend=0.01), no significant difference in the HPV16 integration status was observed between the cases with a punctate signal only and the cases with mixed punctate and diffuse signals (P=0.4). In conclusion, ISH using the Inform HPV III probe seems comparable to PCR for detecting HPV DNA in cervical tissue with CINs. False-negative ISH results appear to be associated with the lower copy numbers of the episomal HPV16 but not with the ability of the Inform HPV III probe to detect specific HPV types. In addition, signal patterns, especially a mixed punctate and diffuse pattern of HPV, cannot be reliably used to predict viral integration status.