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	肝炎基因诊断芯片对丙型肝炎病毒基因分型检测的初步研究
其他题名	Genotyping of hepatitis C virus by hepatitis gene diagnosis microarray
	赵伟 1; 刘伟 1; 刘全俊 2; 张林 1; 周镇先 1; 刘新珏 1; 张汉荣 1
来源期刊	中华医学杂志
ISSN	0376-2491
出版年	2002
卷号	82 期号:18 页码:1249-1253
中文摘要	目的研究丙型肝炎病毒（HCV）基因分型诊断芯片的制备和其对丙型肝炎患者血清HCVRNA基因分型诊断的准确性。方法根据丙型肝炎病毒5′端非编码区末端（5UTR）和C区设计引物和特异性探针，将设计的20条特异性寡核苷酸探针用核苷酸合成仪合成后，配制成浓度为50mumol/ml的点样液，用点样仪点到特殊处理的玻片介质上，制成丙型肝炎病毒基因分型诊断芯片，收集HCVRNA阳性的丙型肝炎患者血清60份作为实验组，健康人血清60份作为对照组，用荧光定量PCR仪对两组血清进行HCVRNA定量检测，实验组血清HCVRNA含量均大于500拷贝/ml，对照组血清HCVRNA含量均小地500拷贝/ml，两组血清进行HCVRNA分离纯化，逆转录反应，巢式PCR扩增后，分别用基因芯片，HCVRNA核酸序列测定法（美国Li-Cor核酸序列测定仪）进行双盲丙型肝炎基因分型检测。结果实验组血清基因诊断芯片检测均为阳性；基因分型；1b型46例，3a型3例。3b型3例，2a型2例，2b型2例，混合型1b+2a2例。1b+3b型2例；核酸序列测定分型1b型50例，3a型3例，3b型3例，2a型2例，2b型2例；基因诊断芯片检测和核酸序列测定检测的基因分型结果符合率为93.3。对照组血清基因芯片检测匀阴性。结论肝炎病毒基因诊断芯片可以用于检测血清HCVRNA，并进行基因诊断分型，准确率较率。
英文摘要	Objective To study the preparation of hepatitis C virus (HCV) diagnosis microarray and its accuracy in diagnosis of gene type of hepatitis C virus. Methods Probe and primer and primers were designed in 5’-untranslated region and C region of hepatitis C virus gene. The probes were synthesized by DNA synthesizer. Solutions of probe of the final concentration of 50 mumol/L were made by dissolving the probes into sodium carbonate buffer. Hepatitis C virus genotype array spotting was performed by pin-based spotting robot PixSys55OO with CMP3 pin. The gene chips were prepared by spotting the probes onto the specially treated glass sliders. Sixty HCV RNA positive serum samples were obtained from the in-patients of the Nanjing Second Hospital (experimental group), and 60 HCV RNA negative serum samples were obtained from the healthy people undergoing physical examination (control group). Quantitative examination of serum HCV RNA was made by fluorescent quantitation PCR. The HCV RNA in the serum specimens of the experimental group (with the HCV RNA concentration of more than 500 copies/ml) and of the control group (with the HCV RNA concentration of less than 500 copies/ml) was isolated and purified, underwent reversed transcription and nested PCR to be amplified, and then genotyped by gene microarray and HCV RNA sequencing. During the experiment, double blind method was used. Results Tested by the gene microarray, the serum specimens in the experimental group were all HCV RNA positive, out of which 46 cases were lb type, 3 cases were 3a type, 3 cases were 3b type, 2 cases were 2a type, 2 eases were 2b type, 2 cases were lb + 2a type, and 2 cases were 3a type. Tested by nucleotide sequencing assay, 50 cases were lb type, 3 cases were 3a type, 3 cases were 3b type, 2 oases were 2a type, and 2 cases were 2b type. The double-blind test results showed a coincidence rate of 93.3% in genotyping HCV by these two methods. Conclusion Hepatitis gene microarray can be used in detection of serum HCV RNA and in diagnostic genotyping with great accuracy.
中文关键词	C型肝炎样病毒属 ; 基因型 ; 基因芯片 ; 丙型肝炎 ; 诊断
英文关键词	hepatitis C-Like viruses genotype
语种	中文
国家	中国
收录类别	CSCD
WOS类目	MEDICINE GENERAL INTERNAL
WOS研究方向	General & Internal Medicine
CSCD记录号	CSCD:1089081
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/203751
作者单位	1.东南大学医学院附属南京市第二医院, 南京, 江苏 210003, 中国; 2.东南大学, 分子与生物电了国家重点实验室
推荐引用方式 GB/T 7714	赵伟,刘伟,刘全俊,等. 肝炎基因诊断芯片对丙型肝炎病毒基因分型检测的初步研究[J],2002,82(18):1249-1253.
APA	赵伟.,刘伟.,刘全俊.,张林.,周镇先.,...&张汉荣.(2002).肝炎基因诊断芯片对丙型肝炎病毒基因分型检测的初步研究.中华医学杂志,82(18),1249-1253.
MLA	赵伟,et al."肝炎基因诊断芯片对丙型肝炎病毒基因分型检测的初步研究".中华医学杂志 82.18(2002):1249-1253.