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运用限制性片段长度多态性技术分析水痘患者中水痘-带状疱疹病毒ORF38和ORF54的酶切消化模式

Enzymatic Digestion Pattern of Varicella Zoster Virus ORF38 and ORF54 in Chickenpox Patients Using RFLP Technique.

作者信息

Safarnezhad Tameshkel Fahimeh, Karbalaie Niya Mohammad Hadi, Keyvani Hossein

机构信息

Gastrointestinal & Liver Disease Research Center (GILDRC), Firuzgar Hospital, Tehran, Iran.

Dept. of Virology, Iran University of Medical Sciences, Tehran, Iran.

出版信息

Iran J Pathol. 2016 Winter;11(1):35-40.

Abstract

BACKGROUND

Varicella zoster virus (VZV) causes chickenpox in children and zoster (zona) in the elderly. Using RFLP-PCR method for detection of VZV specific SNPs ORF38, 54 and 62 could distinguish the profile of VZV isolates. The aim of this study was to investigate enzymatic digestion pattern of VZV ORF38 and ORF54 in chickenpox patients using RFLP technique.

METHODS

Thirty-eight chickenpox patients, who referred to the hospitals of Iran University of Medical Sciences in Tehran from May 2010 to June 2015 were enrolled in this cross sectional study. After the DNA extraction, PCR amplification of 38 VZV isolates performed by specific primers of ORFs 38 and 54, then RFLP assay and digestion carried out by PstI (for ORF38) and BglI (for ORF54) restriction enzymes.

RESULTS

Of 38 positive VZV DNA, the mean age (yr)±SD was 34.4±23.3 (range: 7-89). 22 (57.9%) were female and 16 (42.1%) were male. The predominant VZV profile of BglI(+) PstI(+) were 89.5% (34/38) followed by 10.5% (4/38) PstI(+) BglI‾. Statistical analysis showed that there was no significant relationship between genotype, age, sex, and year of infection variables (P value> 0.05). The common VZV genotype among Iranian patients with chickenpox and zona infection is genotype BglI(+) PstI(+) followed by PstI(+) BglI‾.

CONCLUSION

There are different VZV circulating genotypes that call for for more research on this field by widely population and other methods such as nucleotide sequencing to justify the accurate VZV genotype prevalence in Iran.

摘要

背景

水痘带状疱疹病毒(VZV)可引起儿童水痘和老年人带状疱疹(缠腰龙)。使用RFLP-PCR方法检测VZV特异性单核苷酸多态性(SNP)ORF38、54和62可区分VZV分离株的特征。本研究的目的是使用RFLP技术研究水痘患者中VZV ORF38和ORF54的酶切模式。

方法

本横断面研究纳入了2010年5月至2015年6月转诊至德黑兰伊朗医科大学附属医院的38例水痘患者。DNA提取后,使用ORF 38和54的特异性引物对38株VZV分离株进行PCR扩增,然后进行RFLP分析,并用PstI(用于ORF38)和BglI(用于ORF54)限制性内切酶进行酶切。

结果

在38份VZV DNA阳性样本中,平均年龄(岁)±标准差为34.4±23.3(范围:7 - 89岁)。女性22例(57.9%),男性16例(42.1%)。BglI(+) PstI(+)的主要VZV特征占89.5%(34/38),其次是10.5%(4/38)的PstI(+) BglI‾。统计分析表明,基因型、年龄、性别和感染年份变量之间无显著关系(P值>0.05)。伊朗水痘和带状疱疹感染患者中常见的VZV基因型是BglI(+) PstI(+)基因型,其次是PstI(+) BglI‾。

结论

存在不同的VZV流行基因型,这需要通过更广泛的人群以及其他方法(如核苷酸测序)对该领域进行更多研究,以确定伊朗VZV基因型的准确流行情况。

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