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[膜式DNA芯片法男性人乳头瘤病毒基因分型的研发与临床应用]

[Development and clinical application of male human papillomavirus genotyping by membrane DNA chip].

作者信息

Jin Yu-Ji, Yue Li-Jie, Tao Lin, Deng Fang-Mei, Li De-Fa, He Wen-Shan, Liu Jie, Xiang Jian, Cai Hong, Tu Qian-Qian, Hong Min

机构信息

Department of Clinical Laboratory, Shenzhen Children's Hospital, Shenzhen, Guangdong 518026, China.

出版信息

Zhonghua Nan Ke Xue. 2008 Feb;14(2):106-9.

Abstract

OBJECTIVE

To develop a new method for the detection of male human papillomavirus (HPV) genotypes and to investigate its clinical application value.

METHODS

With computer assistance and based on the classical common primers MY09/11, modified PGMY09/11 with 23 HPV subtypes for PCR and Genbank data on HPV, we designed probes for the simultaneous detection of 18 high-risk subtypes (HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 83 and MM4) and 5 low-risk subtypes (HPV-6, 11, 42, 43 and 44) and fixed them to the special membrane to make a DNA chip. A total of 112 male urethral samples were collected with swabs and studied for the clinical value. Meanwhile the single subtypes of HPV positive were sequenced and the standard samples detected for their sensitivity.

RESULTS

Of the total number, 25 samples were found to be HPV positive, 13 single HPV infection and 12 multiple infection. Nine HPV gene subtypes were noted in the samples: 6, 11, 16, 18, 33, 35, 43, 56 and 73, with sensitivity up to 10 copies of HPV DNA.

CONCLUSION

Human papillomavirus genotyping by the membrane DNA chip is applicable to the diagnosis of male HPV infection as well as to the related epidemic and etiological investigation.

摘要

目的

开发一种检测男性人乳头瘤病毒(HPV)基因型的新方法,并探讨其临床应用价值。

方法

在计算机辅助下,基于经典通用引物MY09/11,设计针对23种HPV亚型的改良PGMY09/11用于PCR,并依据HPV的Genbank数据,设计用于同时检测18种高危亚型(HPV-16、18、31、33、35、39、45、51、52、53、56、58、59、66、68、73、83和MM4)和5种低危亚型(HPV-6、11、42、43和44)的探针,并将其固定于特制膜上制成DNA芯片。用拭子共采集112份男性尿道样本,研究其临床价值。同时对HPV阳性的单一亚型进行测序,并检测标准样本的敏感性。

结果

在全部样本中,发现25份HPV阳性,其中13份为单一HPV感染,12份为多重感染。样本中检测到9种HPV基因亚型:6、11、16、18、33、35、43、56和73,对HPV DNA的敏感性高达10拷贝。

结论

膜DNA芯片进行人乳头瘤病毒基因分型适用于男性HPV感染的诊断以及相关流行病学和病因学调查。

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