摘要
目的建立人乳头瘤病毒(Human papillomavirus,HPV)基因分型芯片技术,采用基因测序技术对该芯片进行评价,探讨其临床使用价值。方法利用基因工具软件比对HPVL1基因,设计可扩增高危型HPV的通用简并引物和可以鉴别HPV型别的基因探针,仪器将探针点样于尼龙膜上,制备HPV分型基因芯片。芯片检测的样本结果与HPV基因测序的结果进行比较,以评价芯片的分析性能。结果建立的HPV基因芯片的灵敏度为102cfu/ml,重复性检测的HPV型符合率为96.7%(87/90),与基因测序结果分型的符合率高达94.3%(66/70),可以检测常见的高危型和导致性传播疾病的HPV型别,可在6～7h内出结果,结果可通过仪器判读。结论HPV基因芯片具有较高的敏感性和重复性,与基因测序的结果符合率高,具有临床推广使用潜力。
Objective To establish and evaluate a gene microarray for HPV detection and typing. Methods Primers for amplifying HPV L1 gene and probes for differentiating HPV genotypes were designed with Primer 5.0. Probes were arrayed by non-contact dotting machine. HPV L1 fragments were cloned and transinfected into E coli DH5a. HPV L1 was sequenced to evaluate the HPV genotyping chip＇s performance. Results The sensitivity and reproducibility of the microarray＇s detection were 102CFU/mL and 96.7%（87/90） respectively. The coincidence rate between chip determination and sequencing was 94.3%（66/70）. The chip was capable of identifying 14 high risk HPV types for cervical carcinoma and HPV-6 and -11 for warts. The bench time from DNA extraction to identification by chip reader was around 6 hours. Conclsuion The microarray we developed was sensitive and specific for HPV typing and had high accordance rate with sequencing results. It will be useful tool to early and rapidly determine HPV infection in clinical laboratories.
出处
《中国热带医学》
CAS
2009年第6期990-993,共4页
China Tropical Medicine
基金
海南省自然科学基金资助项目(No:30647)
