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茅海燕, 卢亦愚, 陈寅, 李榛, 徐昌平, 张严峻, 严菊英. 腺病毒荧光定量PCR快速检测方法建立[J]. 中国公共卫生, 2010, 26(1): 95-96. DOI: 10.11847/zgggws2010-26-01-51
引用本文: 茅海燕, 卢亦愚, 陈寅, 李榛, 徐昌平, 张严峻, 严菊英. 腺病毒荧光定量PCR快速检测方法建立[J]. 中国公共卫生, 2010, 26(1): 95-96. DOI: 10.11847/zgggws2010-26-01-51
MAO Hai-yan, LU Yi-yu, CHEN Yin, . Development of SYBR Green real-time PCR assay for detection of adenovirus[J]. Chinese Journal of Public Health, 2010, 26(1): 95-96. DOI: 10.11847/zgggws2010-26-01-51
Citation: MAO Hai-yan, LU Yi-yu, CHEN Yin, . Development of SYBR Green real-time PCR assay for detection of adenovirus[J]. Chinese Journal of Public Health, 2010, 26(1): 95-96. DOI: 10.11847/zgggws2010-26-01-51

腺病毒荧光定量PCR快速检测方法建立

Development of SYBR Green real-time PCR assay for detection of adenovirus

  • 摘要: 目的 建立腺病毒双链嵌合荧光染色(SYBR Green)实时定量PCR快速检测方法,用于腺病毒感染的早期诊断及病毒核酸定量分析。方法 建立并优化SYBR Green荧光定量PCR反应体系和反应条件,评价该方法的特异性、灵敏度和重复性,同时进行熔解曲线分析,构建定量分析模型,并对84份临床样本进行检测。结果 该方法与其他呼吸道病毒无交叉反应,检测灵敏度102拷贝/μL,熔解曲线显示单一的峰,熔解温度为(84.9±0.1)℃,临床样本检测阳性率达30.95%。结论 建立了快速、敏感、特异的腺病毒SYSR Green荧光定量PCR检测方法,适用于腺病毒的早期快速诊断。

     

    Abstract: Objective To develop a rapid SYBR Green real-tmie PCR a ssay for early detection of human adenovirus and quantification of the virus nucleic acid.Methods The amplification system and the rmal cycle condition of the SYBR Green real-tmie PCR were developed and optmiized.Specificity,sensitivity and reproducibility of the method were evaluated.The characteristic Tm of the target was determinded by preforming a melting curve analysis on the PCR product.The standard curve and virus quantitative analysis model were constructed.Eighty-fourc linical specmiens were detected with the method.Results No cross-amplification was observed when other respiratory virus was tested.As few as 102 copies/μl plasmids were reliably detected by the method.Melting curve analysis showed a single melting peak with a Tm of 84.9±0.1℃.A deno virus DNA was detected in 30.95% of the clinical specmiens.Conclusion Arapid,sensitive and specific SYBR Green real-tmie PCR for analysis of adenovirus DNA was established which was suitable for early diagnosis and virus quantification.

     

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