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The application of alkaline phosphatase labeled hbv probe in serum detection
Tu, ZM; Ke, LH; He, GY
刊名Virus genes
2004-03-01
卷号28期号:2页码:151-156
关键词Alkaline phosphatase Cross-linking Hbv probe Hepatitis Non-radioactive Serum detection Virus dna
ISSN号0920-8569
通讯作者Tu, zm(tuzhiming9@sina.com)
英文摘要A method using non-radioactive material alkaline phosphatase to label hbv dna as probe has been studied and used in clinical experiments to detect the hbv dna in hepatitis serum. alkaline phosphatase coupled with polyethyleneimine (pei) using p-benzoquine as cross-linking reagent. the modified phosphatase was covalently linked to single strand dna using glutraldehyde. such single strand dna enzyme complexes have been tested for blot hybridization, after hybridization and incubation with a substrate solution, sequences complementary to the probe can be visualized directly in 1 h. the minimum amount about 10 pg of target dna has been detected in this way, 32 p labeled probes are autoradiography 1 h after hybridization can only detect 10 ng, so the enzyme labeled probe is more sensitive than isotope labeled probe in 1 h fast test. comparing the enzyme-labeled hbv dna probe with 32 p labeled the same one, positive proportion of detecting the hbv dna in hepatitis patients was about 95.7%. because the positive patient's serum detected by 32 p labeled probe were selected through 1-week radiation, alkaline phosphatase labeled probes are color developed for only 1 h. our experiment certified that it is a sensitive, specific, easy, rapid, safe and economical probe labeling and clinical virus dna detection method.
WOS关键词INSITU HYBRIDIZATION ; BLOT HYBRIDIZATION ; DNA PROBES ; COLORIMETRIC METHOD ; NUCLEIC-ACIDS ; BIOTIN ; RNA ; QUANTITIES ; SEQUENCES ; ASSAY
WOS研究方向Genetics & Heredity ; Virology
WOS类目Genetics & Heredity ; Virology
语种英语
出版者KLUWER ACADEMIC PUBL
WOS记录号WOS:000189075400001
内容类型期刊论文
URI标识http://www.corc.org.cn/handle/1471x/2375187
专题武汉病毒研究所
通讯作者Tu, ZM
作者单位1.Huazhong Univ Sci & Technol, Sch Life Sci & Technol, China UK HUST RRes Genet Engn & Gen Joint Lab, Wuhan 430074, Peoples R China
2.Acad Sinica, Wuhan Inst Virol, Wuhan 430071, Peoples R China
3.Huazhong Univ Sci & Technol, Wuhan 430074, Peoples R China
推荐引用方式
GB/T 7714
Tu, ZM,Ke, LH,He, GY. The application of alkaline phosphatase labeled hbv probe in serum detection[J]. Virus genes,2004,28(2):151-156.
APA Tu, ZM,Ke, LH,&He, GY.(2004).The application of alkaline phosphatase labeled hbv probe in serum detection.Virus genes,28(2),151-156.
MLA Tu, ZM,et al."The application of alkaline phosphatase labeled hbv probe in serum detection".Virus genes 28.2(2004):151-156.
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