期刊文献+

实时定量RT-PCR检测博尔纳病病毒方法的建立 被引量:2

Establishment of real-time RT-PCR for detection of Borna disease virus RNA
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摘要 目的建立检测博尔纳病病毒(BDV)RNA的实时定量RT-PCR(real-time RT-PCR)方法。方法设计BDV-p40基因特异性引物和探针,优化反应条件;并以已知的BDV-p40基因扩增产物的克隆为标准品,建立标准曲线;然后对已知BDV RNA进行实时定量RT-PCR的特异性和敏感性检测。结果建立了特异检测BDV RNA的实时定量RT-PCR技术,并且可以检测到感染细胞中1个拷贝以上BDV RNA。结论本技术可以直接用于BDV感染的检测和定量分析。 Objective To establish the real-time RT-PCR method for detection of Boma disease vires (BDV) RNA. Methods The specific primers and the fluorescence probe of BDV-p40 were designed and synthesized; PCR products of BDV-p40 mRNA were cloned and used as standard template to develop the standard curve for samples detection;meanwhile the specificity and sensitivity of the method were performed. Results The realtime RT-PCR method for detection of BDV RNA was established and it could detect the total RNA of BDV infection cells which contained more than one copy of BDV genome. Conclusion The real-time RT-PCR method can be used to detect BDV infection and quantitatively analyse.
出处 《哈尔滨医科大学学报》 CAS 北大核心 2007年第4期293-295,共3页 Journal of Harbin Medical University
基金 国家自然科学基金资助项目(30371242) 黑龙江省杰出青年科学基金资助项目(JC04-05)
关键词 博尔纳病病毒 逆转录聚合酶链反应 实时PCR Boma disease vires reverse-transcription PCR real-time PCR
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参考文献12

  • 1Chalmers RM,Thomas DR,Salmon RL.Borna disease virus and the evidence for human pathogenicity:a systematic review[J].QJM,2005,98(4):255-274.
  • 2李永杰,王得新,张凤民,刘占东,杨爱英,生田和良.中国慢性疲劳综合征患者血浆中BDV-p24抗体的检测[J].中华实验和临床病毒学杂志,2003,17(4):330-333. 被引量:13
  • 3李桂梅,马培林,张凤民,林玉奇,谷鸿喜,生田和良.用巢式RT-PCR检测精神分裂症患者血液标本中博尔纳病病毒p24基因[J].中国微生态学杂志,2003,15(2):71-72. 被引量:7
  • 4Miranda HC,Nunes SO,Calvo ES,et al.Detection of Borna disease virus p24 RNA in peripheral blood cells from Brazilian mood and psychotic disorder patients[J].J Affect Disord,2006,90(1):43-47.
  • 5李小光,张凤民,马培林,陈小贝,杨爱英.柯萨奇B组病毒的RACE扩增与序列分析[J].中国地方病学杂志,2005,24(2):161-163. 被引量:2
  • 6Watanabe M,Lee BJ,Kamitani W,et al.Neurological diseases and viral dynamics in the brains of neonatally borna disease virus-infected gerbils[J].Virology,2001,282(1):65-76.
  • 7Livak KJ,Flood SJ,Marmaro J,et al.Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization[J].PCR Methods Appl,1995,4(6):357-362.
  • 8Heid CA,Stevens J,Livak KJ,et al.Real time quantitative PCR[J].Genome Res,1996,6(10):986-994.
  • 9Gibson UE,Held CA,Williams PM.A novel method for real time quantitative RT-PCR[J].Genome Res,1996,6(10):995-1001.
  • 10徐平,谢鹏,邹德智,左联.博尔纳病病毒荧光定量套式RT-PCR检测方法的建立[J].重庆医科大学学报,2003,28(6):688-691. 被引量:15

二级参考文献25

  • 1曲波,刘立志,于维汉,王彦民.柯萨奇3m病毒和T-2毒素致小鼠慢性心肌损伤细胞免疫功能改变[J].中国地方病学杂志,2004,23(1):36-38. 被引量:1
  • 2Rott R. Becht H. Natural and experimental Borna disease in animals[J]. Curt Top Microbiol Immunol, 1995,190 : 17-30.
  • 3Rott R, Herzog S-, Fleischer B, et al. Detection of serum antibodies to Borna disease virus in patients with psychiatric disorders[J]. Science, 1985,755-756.
  • 4Sauder C, Muller A,Cubitt B, et al. Detection of Borna disease virus (BDV) antibodies and BDV RNA in paychiatric patients:evidence for high sequence conservation of human blood-derived BDV RNA[J]. Journal of Virology, 1996,7713-7724.
  • 5Nakamura Y,Nakaya T,Hagiwara K,et al. High susceptibility of Mongolian gerbil (Meriones unguiculatus)to Borna disease virus[J]. Vaccine, 1999,17 (5) : 480-489.
  • 6Qiushui He, Nhu Nguyen. Tran Minh, et al. Cytokinem RNA Expression and Proliferative Responses Induced by Pertussis Toxin, Filamentous Hemagglutinin, and Pertactin of Bordetella pertussis in the Peripheral Blood Mononuclear Cells of Infected and Immunized schoolchildren and Adults[J]. Infect Immun,1998,66 (8):3796-3801.
  • 7Tsuji K,Toyomasu K,Imamura Y,et al. No association of borna disease virus with psychiatric disorders among patients in northern Kyushu[J]. Japan J Med Virol, 2000,61(3): 336-340.
  • 8Kishi M, Nakaya T, Nakamura Y, et al. Prevalence of Borna disease virus RNA in peripheral blood mononuclear cells from blood donors[J]. Med Microbiol Immunol (Berl), 1995,184 (3): 135-138.
  • 9Iizuka N, Kuge S, Nomoto A .Complete nucleotide sequence of the genome of coxsackievirus B1 [J]. Virology , 1988,2: 64-73.
  • 10Polacek C, Lundgren A, Andersson A. Genomic and phylogenetic characterization of coxsackievirus B2 prototype strain Ohio-1 [J].Virus Res, 1999,59(2) :229-238.

共引文献31

同被引文献40

  • 1马培林,张凤民,李桂梅,杨爱英,李懿宏,谷鸿喜,生田和良.博尔纳病病毒自然感染状况及其核苷酸序列[J].中国公共卫生,2004,20(4):408-410. 被引量:8
  • 2王振海,谢鹏,韩玉霞,詹军.宁夏及其周边地区博尔纳病病毒感染的分子流行病学研究[J].中华流行病学杂志,2006,27(6):479-482. 被引量:20
  • 3赵立波,谢鹏,李亚军,牟君,刘庆军,周锡国,杨建,吴小容.精神分裂症患者血液标本检测博尔纳病病毒核酸[J].中华微生物学和免疫学杂志,2006,26(12):1101-1101. 被引量:6
  • 4赵立波,谢鹏,牟君,李亚军,邹德智,刘庆军,周锡国,杨建.重庆地区抑郁症患者博尔纳病病毒感染的分子生物学研究(英文)[J].中国神经精神疾病杂志,2007,33(1):18-22. 被引量:6
  • 5Nunes SO, Itano EN, Amarante MK, et al. RNA from Borna disease virus in patients with schizophrenia, schizoaffective patients, and in their biological relatives. J Clin Lab Anal, 2008, 22(4) : 314 -320.
  • 6Bode L, Detlef E, Ludwig HD. Biology of depression. Published online. New York: Wiley, 2008: 583-615.
  • 7Patti AM, Vulcano A, Candelori E, et al. Serological evidence for Borna disease virus infection in children, cats and horses in Sicily (Italy). APMIS Suppl, 2008,124: 77-79.
  • 8Hagiwara K, Asakawa M, Liao L, et al. Seroprevalence of Boma disease virus in domestic animals in Xinjiang, China. Vet Microbiol, 2001, 80(4) : 383-389.
  • 9Pletnikov MV, Rubin SA, Vogel MW, et al. Effects of genetic background on neonatal Borna disease virus infection-induced neurodevelopmental damage. II. Neurochemical alterations and responses to pharmacological treatments. Brain Res, 2002, 944 (1-2) : 108-123.
  • 10周爱儒,查锡良生物化学.6版.北京:人民卫生出版社,2005:43-47.

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