發現潛在性高風險人類乳突病毒感染之基因型和細胞學異常的關聯性

Discovery of Potential High Risk HPV Genotypes Associated with Abnormal Cytology

楊淑理(Shu-Li Yang) ； 曹國倩(Kuo-Chien Tsao) ； 黃瓊瑰(Chung-Guei Huang) ； 楊翾(Shuan Yang) ； 林亞筑(Ya-Jhu Lin) ； 施信如(Shin-Ru Shih)

《Journal of Biomedical & Laboratory Sciences》 25卷2期 (2013/06) Pp. 70-76

摘要

人類乳突病毒（HPV）感染是發生子宮頸細胞病變的元兇，目前市售HPV基因分型檢測套組僅針對常見的基因型設計，未能涵蓋所有的基因型，而這些未被檢測出來的基因型引起子宮頸細胞病變所扮演的角色和重要性未適切的評估。本研究中，我們收集2010年2月至2011年5月間，至林口長庚醫院婦產科門診就診的婦女，進行HPV基因分型檢測的子宮頸拭子檢體共4333例，其中51例（1.2%）以HPV基因分型檢測套組無法鑑定基因型的個案。進一步以基因定序分析，結果顯示有19.6%（10/51）屬於高風險基因型，包括了HPV-51，HPV-52，HPV-68和HPV isolate 06，其中HPV-52還發現2種不同的核酸變異。除此之外，還包括了3例低風險基因型、34例風險未確定基因型、2例未定義基因型及2例未知基因型的HPV感染。而造成這些案例無法分型的主要原因包括部分位點基因序列不同無法與探針進行雜交反應，及感染的基因型不在分型檢測範圍。這些案例中有16/51例（31.4%）合併子宮頸細胞異常，持續追蹤個案細胞學結果也顯示，有5/31例（16.1%）的個案會進展為更差的細胞學異常。現行HPV基因分型檢測套組已能涵括盛行率較高的HPV基因型，但本研究結果顯示，HPV-30，HPV-87，HPV-90，HPV-91等基因型的感染都會引起細胞異常，應屬於潛在性的高風險基因型，這些基因型因為一般HPV檢測套組無法檢測出來而被忽略，及核酸的變異或多樣性無法檢測出這些潛在性高風險基因型。我們建議使用基因分型檢測套組的實驗室，針對HPV PCR出現增幅產物但無法被明確分型的個案，可進一步進行基因序列分析，以提供正確完整的檢驗資訊，供臨床醫師診斷參考。

關鍵字

人類乳突病毒 ； 高風險基因型 ； 低風險基因型 ； 基因分型檢測套組 ； 基因定序

並列摘要

Human papillomavirus (HPV) is the most important risk factor for the development of cervical cancer. A common problem is the failure to cover all genotypes for commercially available HPV genotyping test kits. Those rare or undefined genotypes have not been detected that cause cervical dysplasia of the role and importance not been appropriate assessment. Between February 2010 and May 2011, a total of 4333 specimens were subjected to our laboratory for the genotyping analysis. In those 4333 cases, 51 cases (1.2%) could not been identified by HPV genotyping test kits. Further gene sequencing results showed that 19.6% (10/51) of which are high-risk genotype, including HPV-51, HPV-52, HPV-68 and HPV isolate 06. We also found that there are two different nucleic acid variants in HPV-52. In addition, 3 cases of low-risk genotypes and 34 cases with risk undefined genotypes, 2 cases of undetermined and 2 cases of unknown genotype HPV infection were identified. The reasons why those 51 cases could not be typed are DNA mismatches between the detection probe and target sequences or genotypes are not covered by the kit. In those 51 cases, 16 cases (31.4%) merger abnormal cervical cells, and keep track of cytology results also showed 16.1% (5/31) of cases will progress worse cytological abnormalities. Our results indicated that HPV-30, HPV-87, HPV-90, HPV-91 infections cases, are high-risk genotypes which are ignored by the test kit because of the nucleic acid variation or diversity. In order to provide the clinicians with precise and correct laboratory examination results, a further sequence analysis is recommended for cases that produced positive results in the HPV PCR but negative in the subsequent hybridization procedure when using the HPV commercial kit.

並列關鍵字

Human papillomavirus ； high-risk genotype ； low-risk genotype ； HPV genotyping test kits ； gene sequencing