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首页> 外文期刊>AIDS Research and Human Retroviruses >Short Communication: Serum-Based Assay Accurately Detects Single Nucleotide Polymorphisms of IL28B and SOCS3 in HIV/Hepatitis C Virus-Coinfected Subjects
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Short Communication: Serum-Based Assay Accurately Detects Single Nucleotide Polymorphisms of IL28B and SOCS3 in HIV/Hepatitis C Virus-Coinfected Subjects

机译：简短交流：基于血清的分析可准确检测HIV /丙型肝炎病毒合并感染受试者中IL28B和SOCS3的单核苷酸多态性

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摘要

Single nucleotide polymorphisms (SNPs) have become important in predicting treatment response to interferon containing anti-hepatitis C virus (HCV) therapy in HCV and HIV/HCV-infected patients. A reliable method for extracting host DNA from serum for genotyping assays would present a practical alternative for clinicians and investigators seeking to perform SNP analyses in HCV-infected patients, particularly in resource-limited settings. Human genomic DNA was extracted from peripheral blood mononuclear cells (PBMCs) and serum of 51 HIV/HCV coinfected patients using the QIAamp DNA Blood Mini Kit and QIAamp Min Elute Virus Spin Kit, respectively. Genotyping assays for the IL28B SNP (rsl2979860) and SOCS3 SNP (rs4969170) were performed using the commercially available ABI Taqman allelic discrimination kit and reverse transcriptase-polymerase chain reaction (RT-PCR) was performed using 50 cycles. Results of the genotyping assays using DNA from both PBMCs and cell-free serum were determined separately and then analyzed for concurrence. Genotype analyses performed using DNA isolated from PBMCs or cell-free serum showed a 100% agreement between the IL28B genotyping results from the serum and PBMC isolates and 98% agreement for SOCS3 SNP. This novel serum-based assay to isolate DNA fragments from the serum of HIV/HCV-coinfected subjects can accurately determine a subject's genotype for IL28B (rsl2979860) and SOCS3 (rs4969170). This assay could be immediately valuable for detecting clinically relevant SNPs from serum in cases in which PBMCs are not available.
机译：单核苷酸多态性（SNP）在预测HCV和HIV / HCV感染患者对含干扰素的抗丙型肝炎病毒（HCV）治疗的治疗反应中已变得重要。一种可靠的从血清中提取宿主DNA进行基因分型分析的方法，将为寻求在HCV感染患者中进行SNP分析的临床医生和研究人员提供一种实用的替代方法，特别是在资源有限的环境中。使用QIAamp DNA Blood Mini Kit和QIAamp Min Elute Virus Spin Kit分别从51名HIV / HCV合并感染患者的外周血单核细胞（PBMC）和血清中提取人基因组DNA。使用可商购的ABI Taqman等位基因鉴别试剂盒对IL28B SNP（rs12979860）和SOCS3 SNP（rs4969170）进行基因分型测定，并使用50个循环进行逆转录酶-聚合酶链反应（RT-PCR）。使用来自PBMC和无细胞血清的DNA分别进行基因分型测定的结果，然后分析其并发性。使用从PBMC或无细胞血清中分离的DNA进行的基因型分析显示，血清和PBMC分离物的IL28B基因分型结果之间有100％的一致性，而SOCS3 SNP的一致性为98％。从HIV / HCV合并感染的受试者的血清中分离出DNA片段的这种基于血清的新测定法可以准确确定受试者的IL28B（rs12979860）和SOCS3（rs4969170）的基因型。如果没有PBMC，该检测方法对于从血清中检测临床相关的SNP可能立即有价值。

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