人乳头瘤病毒检测及分型基因芯片制备的初步研究

目的开发研制对人乳头瘤病毒(HPV)进行检测及分型的基因芯片。方法2003年6月至2003年12月郑州大学第二附属医院将人工合成的HPV通用探针,HPV6,11,16,18特异性寡核苷酸片段作为实验组探针,将与HPV没有同源性的植物基因片段作为内参照基因点样于玻片介质,制成HPV检测及分型芯片,将HPV标准品荧光标记后与芯片杂交,用GMS418扫描仪对芯片进行扫描,以HPV标准品与芯片的杂交结果对芯片检测方法的准确性进行评价。结果该芯片的内参照结果能做到绝对阳性和绝对阴性,各HPV标准品的芯片检测结果显示其在芯片上的相应位置为明显的阳性信号,且各型别之间无错配杂交信号。结论本研究制作的HPV检测及分型芯片可以成功用于HPV的检测及分型,可望用于临床。

Preliminary study of human papillomavirus detection and genotyping DNA microarray

Objection To design a HPV-DNA microarray to detect different HPV genotypes. Methods One common oligonucleotide probe to HPV and four specific oligonucleotide probes to their genotypes(HPV6,11,16,18) were dotted on chips. The non-homology sequences of vegetative genes were used as control. 4 HPV plasmids that contained each type of the human papillomavirus L1 genome were used for the evaluation of this method. HPV plasmids amplified and labled with the fluorescence(CY5) by PCR were hybridized with the microarray. After hybridization the chips were scanned by GMS418 Array Scanner. Results The positive and negative controls were all absolutely right. In all cases ,the plasmids showed clear positive signals on their corresponding positions and there was no mismatched signal. Conclusion This HPV DNA microarray can be used for HPV clinical diagnosis,and it has potential use as a screening test.