IRAP-PCR作为筛查鼻粘膜拭子HERV多态性的工具

IF 0.2 Q4 OTORHINOLARYNGOLOGY ENT Updates Pub Date : 2019-08-08 DOI:10.32448/ENTUPDATES.578602
A. Kepekçi, Merve Seda Ibisoglu, S. Yılmaz, Cenk Kıg
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引用次数: 0

摘要

目的:利用逆转录转座子间多态性聚合酶链反应(IRAP-PCR)技术,通过扩增植物基因组中两个逆转录转座子之间的DNA片段,检测插入多态性。然而,迄今为止,这种方法还没有被报道用于分析人体样本。最近,人类内源性逆转录病毒(HERV)多态性由于其对某些疾病的病理生理学的潜在影响而引起了人们的兴趣。然而,HERV多态性与发展为鼻息肉病(NP)的风险之间的关系尚未得到研究。在本研究中,我们旨在研究是否可以在鼻拭子样本中进行IRAP-PCR,以比较不同鼻粘膜样本中的HERV多态性。方法:对16例鼻拭子标本进行DNA分离。将这些DNA样品用作HERV-K6、HERV-K11、HERV-L1和HERV-L2的IRAP PCR的模板,并通过琼脂糖凝胶电泳分析PCR产物。结果:鼻拭子样本产生了足够的DNA物质,可以成功地进行IRAP-PCR。我们在本研究中测试的四分之三的HERV序列中获得了特定的条带模式。在来自不同患者的样本之间没有检测到多态性。类似地,在从同一患者获得的息肉或鼻粘膜拭子样本之间没有检测到多态性条带。结论:我们首次证明IRAPCR可以在鼻拭子中进行。我们的研究结果表明,这项技术可以作为一种廉价有效的筛查工具,用于研究鼻粘膜疾病与HERV多态性(如鼻息肉病)之间的联系。
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IRAP-PCR As A Tool For Screening HERV Polymorphisms In Nasal Mucosal Swabs
Objective: Inter-retrotransposon polymorphism Polymerase  Chain Reaction (IRAP-PCR) technique allows for  detecting insertional polymorphisms via amplification  of the DNA fragment between two retrotransposons in  plant genomes. However, this method has not been reported  to be used for analyzing human samples to date.  Recently, Human Endogenous Retrovirus (HERV) polymorphisms  gained interest due to their potential effect  on pathophysiology of certain diseases. Nevertheless, the  association between HERV polymorphisms and the risk  for developing nasal polyposis (NP) has not been studied.  In this study, we aimed to investigate whether or not  IRAP-PCR could be performed in nasal swab samples for  comparing HERV polymorphisms in different nasal mucosal  samples. Methods: Nasal swab samples from 16 patients were used  for DNA isolation. These DNA samples were used as templates  for IRAP PCR of HERV-K6, HERV-K11, HERV-L1 and  HERV-L2 and PCR products were analyzed by agarose gel  electrophoresis. Results: Nasal swab samples yielded enough DNA material  for successfully performing IRAP-PCR. We obtained  specific banding patterns the three out of four HERV  sequences tested in this study. No polymorphisms was  detected between samples from different patients. Similarly,  polymorphic bands was not detected between the  polyps or nasal mucosal swab samples obtained from the  same patient. Conclusion: We have, for the first time, shown that IRAPPCR  can be performed in nasal swabs. Our findings suggest  that this technique can serve as an inexpensive and  effective screening tool for investigating links between  nasal mucosal diseases and HERV polymorphisms such as  nasal polyposis.
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来源期刊
ENT Updates
ENT Updates OTORHINOLARYNGOLOGY-
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