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通过对孕妇阴道拭子样本进行宏基因组分析鉴定一种新型人乳头瘤病毒。

Identification of a novel human papillomavirus by metagenomic analysis of vaginal swab samples from pregnant women.

作者信息

Liu Zhijian, Yang Shixing, Wang Yan, Shen Quan, Yang Yan, Deng Xutao, Zhang Wen, Delwart Eric

机构信息

School of Medicine, Jiangsu University, 301 Xuefu Road, Zhenjiang, Jiangsu, 212023, People's Republic of China.

The Fourth Affiliated Hospital of Jiangsu University, 20 Zhengdong Road, Zhenjiang, Jiangsu, 212001, China.

出版信息

Virol J. 2016 Jul 4;13:122. doi: 10.1186/s12985-016-0583-6.

Abstract

BACKGROUND

The number of members in the genus Gammapapillomavirus of Family Papillomaviridae has recently been expanding most rapidly. The aim of this study was to characterize a novel human gammapapillomavirus type identified in a vaginal swab from a 25-year-old pregnant woman suffering from vaginitis.

METHODS

Viral metagenomics method was used to detect the viral sequences in 88 vaginal swab samples collected from 88 pregnant women with vaginitis. A novel papillomavirus, named HPV-ZJ01 (GenBank no. KX082661), was detected in one sample and its complete genome sequence was amplified by PCR and sequenced by Sanger walking. Phylogenetic analyses based on the complete genome and the L1 protein of HPV-ZJ01 and other representative human papillomaviruses were done, respectively. Further PCR screening was performed in vaginal swabs (n = 135), cervical smears (n = 40) and cervical cancer tissues (n = 40) using nested-PCR primers designed based on HPV-ZJ01 sequence to investigate the prevalence of HPV-ZJ01.

RESULTS

The genome of HPV-ZJ01 is 7,358 bp in length with a GC content of 37.8 %. HPV-ZJ01 was predicted to contain six open reading frames (E6, E7, E1, E2, L2, and L1) and a non-coding long control region (LCR). The genome shared the highest overall similarity to HPV-166, with 70.6 % nucleotide sequence identity while its L1 gene shared the highest nucleotide similarity to HPV-162, with 71.1 % sequence identity. Phylogenetic analysis suggested that HPV-ZJ01 belongs to a novel HPV type in the Gamma-PV genus, species Gamma-19, already containing HPV161, HPV162 and HPV166. PCR screening results indicated that none of the other samples were positive for HPV-ZJ01 except the original HPV-ZJ01 positive vaginal swab specimen.

CONCLUSION

The genome sequence of a novel type of species Gamma-19 HPV was characterized. The screening PCR results suggested that HPV-ZJ01 is not associated with any of the cervical cancer samples tested. In order to confirm the prevalence and disease association, if any, for HPV-ZJ01, a further study with different sample types and a larger sample size is needed.

摘要

背景

乳头瘤病毒科丙型乳头瘤病毒属的成员数量最近增长最为迅速。本研究的目的是对从一名患有阴道炎的25岁孕妇的阴道拭子中鉴定出的一种新型人类丙型乳头瘤病毒进行特征分析。

方法

采用病毒宏基因组学方法检测从88名患有阴道炎的孕妇采集的88份阴道拭子样本中的病毒序列。在一个样本中检测到一种名为HPV-ZJ01(GenBank编号:KX082661)的新型乳头瘤病毒,通过PCR扩增其完整基因组序列,并采用桑格步移法进行测序。分别基于HPV-ZJ01和其他代表性人类乳头瘤病毒的完整基因组和L1蛋白进行系统发育分析。使用基于HPV-ZJ01序列设计的巢式PCR引物,对阴道拭子(n = 135)、宫颈涂片(n = 40)和宫颈癌组织(n = 40)进行进一步的PCR筛查,以研究HPV-ZJ01的流行情况。

结果

HPV-ZJ01的基因组长度为7358 bp,GC含量为37.8%。预计HPV-ZJ01包含六个开放阅读框(E6、E7、E1、E2、L2和L1)和一个非编码长控制区(LCR)。该基因组与HPV-166的总体相似性最高,核苷酸序列同一性为70.6%,而其L1基因与HPV-162的核苷酸相似性最高,序列同一性为71.1%。系统发育分析表明,HPV-ZJ01属于丙型乳头瘤病毒属中的一种新型HPV类型,即丙型-19型,该类型已包含HPV161、HPV162和HPV166。PCR筛查结果表明,除了最初的HPV-ZJ01阳性阴道拭子标本外,其他样本均未检测到HPV-ZJ01阳性。

结论

对一种新型丙型-19型HPV的基因组序列进行了特征分析。PCR筛查结果表明,HPV-ZJ01与所检测的任何宫颈癌样本均无关联。为了确认HPV-ZJ01的流行情况及其与疾病的关联(如果有的话),需要使用不同样本类型并扩大样本量进行进一步研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63b6/4943014/192ab63be59b/12985_2016_583_Fig1_HTML.jpg

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