摘要
目的研制人类免疫缺陷病毒(HIV)快速筛查的寡核苷酸芯片并初步应用于临床。方法以HIV2个基因型(包括9个亚型)的32株典型性代表株和HIV2特有的vpx序列的保守序列为靶序列,设计长寡核苷酸探针,并制备成Oligo芯片。样品经随机特异性引物PCR标记后与芯片杂交,PCR产物同时进行测序分析。结果1例HIV患者芯片杂交结果阳性,20名健康对照血清均为阴性。阳性标本的序列分析表明,芯片杂交结果符合测序的分型结果。结论此芯片可初步用于检测血清HIVRNA,并对HIV基因(亚)型进行分析。
Objective To prepare 60 mer oligo microarray chips for detecting human immunodeficiency virus (HIV) and for the clinical application in the detection of AIDS patient. Methods Oligonucleotide probes were designed according to the sequence information of two types of HIV. Oligo microarray was prepared by using Cartesian Microarrayer. Products of the restrictive display PCR were labeled with Cy3. Furthermore, the PCR products were sequenced. Results Using the oligo microarray, HIV infection could be detected in laboratory as well as in clinical assays. Results of hybridization indicated that 1 AIDS patient was positive and 20 health people were negative. The results obtained by sequencing confirmed the results obtained by oligo microarray studies. Conclusion The HIV 60 mer oligo microarray could be used in detecting patient HIV infection and analyzing its genotypes.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2005年第7期616-618,共3页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金资助项目(39880032)
