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[采用巢式聚合酶链反应从急性呼吸道感染婴幼儿中检测人鼻病毒]

[Human rhinovirus detection from infants and young children with acute respiratory infections by nested-polymerase chain reaction].

作者信息

Zhao Lin-qing, Qian Yuan, Zhu Ru-nan, Deng Jie, Wang Fang

机构信息

Laboratory of Virology, Beijing Municipal Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China.

出版信息

Zhonghua Liu Xing Bing Xue Za Zhi. 2006 Feb;27(2):154-6.

Abstract

OBJECTIVE

To develop a rapid, sensitive and specific method for detection human rhinovirus (HRV) from clinical specimens.

METHODS

Primers derived from the highly conserved 5'noncoding region of human rhinovirus were used to develop a nested RT-PCR for detecting HRV. The sensitivity and specificity of the RT-PCR were determined using various RNA while DNA viruses were used as control. Seven hundred and seventy-one specimens collected from children with symptoms of acute respiratory infections from Nov. 2002 to Oct. 2003 were analyzed for HRV by RT-PCR as well as for other respiratory viruses through isolation of virus and indirect immunofluorescent assay.

RESULTS

Only the cDNA from HRV was positive by RT-PCR, indicating the nested RT-PCR was specific. With RT-PCR, HRV were detected in 148 out of 771 specimens (19.2%). As for HRV positive rates, it was found 53.3% in pharyngitis patients; 43.8% in laryngitis patients and 28.7% in bronchitis patients. In Sep. 2002 and from Aug. 2003 to Oct. 2003, HRV positive rates were high (21.6% - 32.6%), with Sep. 2003 in particular--32.6%. From Mar. 2003 to Jul. 2003, HRV positive rates maintained from 16.0% to 19.1%.

CONCLUSION

HRV was one of the important agents for acute respiratory infections in infants and young children in Beijing.

摘要

目的

建立一种从临床标本中快速、灵敏且特异检测人鼻病毒(HRV)的方法。

方法

使用源自人鼻病毒高度保守的5'非编码区的引物,开发用于检测HRV的巢式逆转录聚合酶链反应(RT-PCR)。利用各种RNA测定RT-PCR的敏感性和特异性,同时以DNA病毒作为对照。对2002年11月至2003年10月期间从有急性呼吸道感染症状的儿童中收集的771份标本,通过RT-PCR分析HRV,并通过病毒分离和间接免疫荧光测定法分析其他呼吸道病毒。

结果

RT-PCR仅显示来自HRV的互补DNA(cDNA)呈阳性,表明巢式RT-PCR具有特异性。通过RT-PCR,在771份标本中的148份(19.2%)检测到HRV。至于HRV阳性率,在咽炎患者中为53.3%;在喉炎患者中为43.8%,在支气管炎患者中为28.7%。2002年9月以及2003年8月至10月期间,HRV阳性率较高(21.6% - 32.6%),尤其是2003年9月——为32.6%。2003年3月至7月期间,HRV阳性率维持在16.0%至19.1%之间。

结论

HRV是北京婴幼儿急性呼吸道感染的重要病原体之一。

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