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异源双链迁移率分析在人类免疫缺陷病毒1型基因分型中的应用

[Application of heteroduplex mobility assay in genetic subtyping on human immunodeficiency virus type 1].

作者信息

Chen Liang, Yan Yansheng, Weng Yuwei, Wang Huirong, Wu Shouli, Chen Ge, Zheng Zhaoshuang, Zheng Jian, Yan Pingping

机构信息

Fujian Provincial Sanitary and Anti-Epidemic Station, Department of AIDS Prevention and Control, Fuzhou 350001, China.

出版信息

Zhonghua Liu Xing Bing Xue Za Zhi. 2002 Apr;23(2):127-30.

Abstract

OBJECTIVE

Using heteroduplex mobility assay (HMA) to subtype human immunodeficiency virus type 1 (HIV-1) for the purpose of understanding HIV-1 subtype epidemic in Fujian province.

METHODS

DNA fragments of HIV-1 env gene were amplified from peripheral blood mononuclear cell (PBMC) cocultures of HIV-1 infected individuals by nested polymerase chain reaction (PCR). Heteroduplexs were formed through hybridizing PCR products from the samples and reference plasmid. According to the mobility of heteroduplexs in polyacrylamide gel electrophoresis, HIV-1 subtype from that sample was characterized and further confirmed by nucleotide sequencing analysis.

RESULTS

Thirteen of 15 (86.67%) samples were successfully subtyped by HMA, except 2 failures. Subtype E and B took up 80% (12/15) and 6.67% (1/15) respectively. Results indicated a high concordance between HMA and nucleotide sequencing analysis and concordance rate was 86.67% (13/15).

CONCLUSIONS

Subtype E appeared to be the major epidemic strain of HIV-1 in Fujian. HMA showed the characteristics of fastness, easiness, economic and with high specificity, and can be used in the surviellance for the epidemic strain of HIV-1.

摘要

目的

运用异源双链迁移率分析(HMA)对1型人类免疫缺陷病毒(HIV-1)进行亚型分析,以了解福建省HIV-1亚型的流行情况。

方法

通过巢式聚合酶链反应(PCR)从HIV-1感染个体的外周血单个核细胞(PBMC)共培养物中扩增HIV-1 env基因的DNA片段。将样品的PCR产物与参考质粒杂交形成异源双链。根据异源双链在聚丙烯酰胺凝胶电泳中的迁移率,对该样品的HIV-1亚型进行鉴定,并通过核苷酸测序分析进一步确认。

结果

15个样本中有13个(86.67%)通过HMA成功进行了亚型分析,2个失败。E亚型和B亚型分别占80%(12/15)和6.67%(1/)。结果表明HMA与核苷酸测序分析高度一致,一致率为86.67%(13/15)。

结论

E亚型似乎是福建省HIV-1的主要流行毒株。HMA具有快速、简便、经济且特异性高的特点,可用于HIV-1流行毒株的监测。

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