[1]段晶晶),刘国华),黄学勇),等.柯萨奇病毒B5巢式RTPCR检测方法的建立*[J].郑州大学学报(医学版),2013,(03):349.
 DUAN Jingjing),LIU Guohua),HUANG Xueyong),et al.Establishment of nested RTPCR assay for detection of coxsackie virus B5[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2013,(03):349.
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柯萨奇病毒B5巢式RTPCR检测方法的建立*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2013年03期
页码:
349
栏目:
论著
出版日期:
2013-06-20

文章信息/Info

Title:
Establishment of nested RTPCR assay for detection of coxsackie virus B5
作者:
段晶晶123)刘国华3)黄学勇3)李幸乐3)王芳4)胡小宁13)许汴利13)#
1) 郑州大学公共卫生学院流行病学教研室 郑州 450001
2)郑州市疾病预防控制中心传染病防治科 郑州 450007
3) 河南省疾病预防控制中心传染病预防控制所 郑州 450016
4)郑州市儿童医院感染性疾病科 郑州 450053
Author(s):
DUAN Jingjing123)LIU Guohua3)HUANG Xueyong3)LI Xingle3)WANG Fang4)HU Xiaoning13)XU Bianli13)
1)Department of Epidemiology,College of Public Health, Zhengzhou University,Zhengzhou 450001
2) Department of Infectious Disease Control and Prevention,Zhengzhou Center for Disease Control and Prevention,Zhengzhou 450007
3)Institute of Infectious Disease Control and Prevention,Henan Provincial Center for Disease Control and Prevention,Zhengzhou 450016
4)Department of Infectious Disease,Childrens Hospital of Zhengzhou, Zhengzhou 450053
关键词:
柯萨奇病毒B5巢式RTPCR检测
Keywords:
coxsackie virus B5nested RTPCRdetection
分类号:
R373.2+3
摘要:
摘要目的:建立一种快速、灵敏、特异的柯萨奇病毒B5(CVB5)检测方法。方法:根据GenBank 发表的CVB5河南分离株全基因组序列,在其VP1 区设计并合成巢式RTPCR引物,优化PCR反应条件,建立检测CVB5的巢式RTPCR方法。将已测得TCID50的病毒标准株进行10倍梯度稀释后用上述方法检测,以评价该方法的灵敏性;将5株不同的CVB5毒株和EV71等4株其他肠道病毒毒株及阴性对照用该方法进行扩增,以检测其特异性。用建立的巢式RTPCR方法对52份病毒性脑炎病例样本进行检测。结果:该方法对CVB5的两轮PCR扩增敏感性分别为10 TCID50和10-4 TCID50;所有CVB5毒株用该方法扩增结果均为阳性,所选其他肠道病毒扩增结果均为阴性。52份临床病例样本中检出10份CVB5阳性,阳性率为19.23%。结论:所建立的CVB5巢式RTPCR检测方法具有良好的灵敏度和特异度。
Abstract:
AbstractAim: To establish a rapid,sensitive,and specific method to detect coxsackie virus B5(CVB5).Methods: Two pairs of primers were designed according to the VP1 section of CVB5 isolated from Henan province nucleoprotein gene sequences published in GenBank. With optimized reaction conditions,a reverse transcription nested PCR assay for detection of CVB5 was developed specifically. The standard virus strains which had been identified as TCID50 were serially tenfold diluted and were used to determine the sensitivity of the method,and cDNA extracted from five different CVB5 strains and four other enteroviruses was used as template to assess the specificity of the method.Furthermore,52 samples of viral encephalitis were performed by using this nested RTPCR method.Results: The sensitivity of twowheeled amplification of CVB5 were respectively 10 TCID50 and 10-4TCID50.The amplification results of all CVB5 strains were positive, while the results of four other enteroviruses were negative.Ten positive results were obtained from the 52 samples using this method and the positive rate was 19.23%. Conclusion: A nested RTPCR assay to detect CVB5 has been established successfully.

参考文献/References:

[1] 黄学勇,许玉玲,李幸乐,等. 柯萨奇病毒B5河南分离株全基因组序列测定及分析[J]. 郑州大学学报:医学版,2011,46(6):868
[2] 李华,杨卉娟,柯华昕,等.一株柯萨奇病毒B组5型(Cox.B5)病毒的分离及VP1基因分析[J].医学研究杂志,2011,40(9):55
[3] 胡永峰,赵丽娜,董杰,等.中国萨科奇病毒B5的全基因组测序及其序列分析[J].病毒学报,2010,26(4):283
[4] 王海岩,李岩,徐爱强,等. 柯萨奇B5病毒引起山东省一起无菌性脑膜炎暴发的鉴定及其亲缘进化分析[J].中华流行病学杂志,2010,31(1):64
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[6] 沈建峰,蒋就喜. 病毒性脑炎的分子生物学诊断进展[J]. 医学综述,2010, 16(8):1226
[7] Cheng MF, Chen BC, Huang TS, et al. Clinical application of reversetranscription polymerase chain reaction and intravenous immunoglobulin for enterovirus encephalitis[J]. Jpn J Infect Dis, 2008,61(1):18

相似文献/References:

[1]黄学勇△,许玉玲,李幸乐,等.柯萨奇病毒B5河南分离株全基因组序列测定及分析*[J].郑州大学学报(医学版),2011,(06):868.
 HUANG Xueyong,XU Yuling,LI Xingle,et al.Analysis of genomic characteristics of coxsackie virus B5 strains isolated from Henan province[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2011,(03):868.

备注/Memo

备注/Memo:
*河南省医学科技攻关基金资助重大项目201001015;河南省医学科技攻关计划普通项目201102017;河南省医学科技攻关计划省部共建项目201201003
#通讯作者,男,1958年3月生,主任医师,研究方向:分子流行病学与传染病预防控制, Email: xubl@hncdc.com.cn
更新日期/Last Update: 2013-07-03