异源双链泳动分析法快速检测TT病毒基因型

何忠平; 庄辉; 姚均; 董庆鸣; 戴旺苏; 宋淑静

Abstract

何忠平,庄辉,姚均,董庆鸣,戴旺苏,宋淑静.异源双链泳动分析法快速检测TT病毒基因型[J].Chinese journal of Epidemiology,2003,24(9):801-805

异源双链泳动分析法快速检测TT病毒基因型

Rapid detection of genotypes of TT virus using a heteroduplex mobility assay

Received:August 21, 2002

DOI：

KeyWord: TT病毒异源双链泳动分析基因进化树基因型

English Key Word: TT virus Heteroduplex mobility assay Phylogenetic tree Genotype

FundProject:北京市委组织部优秀人才工程资助项目 (200101)

Author Name	Affiliation
HE Zhong-ping	Department of Microbiology, Peking University Health Science Center, Beijing 100083, China
ZHUANG Hui	Department of Microbiology, Peking University Health Science Center, Beijing 100083, China
YAO Jun	北京地坛医院
DONG Qing-ming	北京地坛医院
DAI Wang-su	北京地坛医院
SONG Shu-jing	北京地坛医院

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Abstract:

目的　建立一种简单、敏感、特异和成本低的TT病毒 (TTV)基因型测定方法。方法采用巢式聚合酶链反应方法 (nPCR)对 96名正常人和 180例各型肝炎患者血清标本进行TTVDNA检测,然后采用异源双链泳动分析法 (HMA)和序列测定分析法对TTVDNA阳性标本进行基因分型。结果　各型病毒性肝炎中TTVDNA阳性率为 2 2.2 % (40 / 180 ),正常人为 19.8% (19/ 96 ),两者差异无显著性 (χ2 =0.2 2 0,P =0.6 39)。在甲、乙、丙、戊型肝炎和非甲～戊型肝炎患者中TTVDNA阳性率分别为2 0.0 % (6 / 30 )、16.7% (5 / 30 )、2 3.3% (7/ 30 )、36.7% (11/ 30 )和 18.3% (11/ 6 0 )。在 4 0例TTVDNA阳性标本中,经HMA法分型,G1型为 5 0.0 % (2 0 / 4 0 ),G2型为 17.5 % (7/ 4 0 ),混合型为 2 5.0 % (10 / 4 0 ),另有 7.5 %(3/ 4 0 )未能分型。 10例TTV混合型感染的标本经克隆测序及基因进化树分析,5例为G1和G2型,2例为G1和G3型,1例为G1和G4型,1例为G2和G3型,1例为G1、G2和G3混合型感染。结论　HMA是一种简单、敏感、特异和经济的分型法,可用于TTV基因型分型。

English Abstract:

Objective To establish a simple, sensitive, specific and less-costly method for detecting genotypes of TT virus (TTV). Methods TTV DNA was tested by nested polymorase chain reaction (nPCR) in sera from 180 patients with different types of viral hepatitis and 96 normal individuals in Beijing. TTV genotypes were determined in 40 sera collected from TTV DNA positive patients by heteroduplex mobility assay (HMA) and through sequencing. Results The positive rates of TTV DNA in viral hepatitis patients and normal individuals were 22.2 %(40/180) and 19.8 %(19/96), respectively ( χ 2= 0.220, P = 0.639 ). TTV DNA positive rates of patients with hepatitis A, B, C, E and non- A to E were 20.0 %( 6/ 30), 16.7 %( 5/ 30), 23.3 %( 7/ 30), 36.7 %( 11/ 30) and 18.3 %( 11/ 60), respectively. Of 40 TTV DNA positive patients, 20( 50.0 %) were TTV G1, 7( 17.5 %) TTV G2,10( 25.0 %) coinfected with different genotypes of TTV, and 3 untyped by HMA. Twenty G1 and 7 G2 detected by HMA were confirmed by sequence analysis. Of 10 patients coinfected with different genotypes of TTV, 5 were G1 and G2, 2 G1 and G3, 1 G1 and G4, 1 G1 and G3, and 1 with G1, G2 and G3 coinfections. Conclusion HMA was recognized as simple, sensitive, specific and less-costly, thus could be used for genotyping of TTV.

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