巢式PCR诊断儿科患者鼻病毒感染的探讨

赵林清; 钱渊; 朱汝南; 邓洁; 王芳

文章摘要

赵林清,钱渊,朱汝南,邓洁,王芳.巢式PCR诊断儿科患者鼻病毒感染的探讨[J].中华流行病学杂志,2006,27(2):154-156

巢式PCR诊断儿科患者鼻病毒感染的探讨

Human rhinovirus detection from infants and young children with acute respiratory infections by nestedpolymerase chain reaction

收稿日期:2005-04-28 出版日期:2014-09-12

DOI：

中文关键词: 鼻病毒巢式逆转录聚合酶链反应

英文关键词: Human rhinovirus Nested reverse transcription polymerase chain reaction

基金项目:北京人类疾病基因诊断基础性研究实验室科研资助项目(JS96004);北京市优秀人才培养专项经费个人资助项目(20042D0300935);北京市科委新星计划资助项目(2004B34)

作者	单位	E-mail
赵林清	100020, 首都儿科研究所病毒研究室北京市感染与免疫中心实验室
钱渊	100020, 首都儿科研究所病毒研究室北京市感染与免疫中心实验室	yqianbjc@263.net
朱汝南	100020, 首都儿科研究所病毒研究室北京市感染与免疫中心实验室
邓洁	100020, 首都儿科研究所病毒研究室北京市感染与免疫中心实验室
王芳	100020, 首都儿科研究所病毒研究室北京市感染与免疫中心实验室

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中文摘要:

目的了解儿科急性呼吸道感染的病毒病原,建立快速、敏感、特异的检测鼻病毒(HRV)的方法。方法根据已发表的HRV的核苷酸序列设计巢式PCR引物,并用多种呼吸道病毒的标准毒株确定方法的特异性。收集2002年11月至2003年10月的急性呼吸道感染患儿标本771例,应用巢式PCR方法检测标本中有无HRV基因片段。结果特异性检测结果显示仅HRVcDNA有预期大小的片段扩增。771例标本中HRV总检出例数为148例(148/771),占19.2%;HRV阳性检出率在咽炎患儿中达到53.3%(8/15),喉炎患儿43.8%(7/16),支气管炎患儿为28.7%(29/101),其他如急性扁桃体炎、急性肺炎等患儿也有较高的HRV阳性检出率。在2002年11月以及2003年8-10月HRV检测阳性率较高(21.6%~32.6%),尤其在2003年9月份HRV检测阳性率最高,达到32.6%;2003年3-7月HRV阳性标本所占百分率较平稳,在16.0%~19.1%之间。结论建立的巢式RT-PCR可以检测到儿科呼吸道感染患儿标本中的鼻病毒基因片段;HRV是北京婴幼儿急性呼吸道感染的重要病毒病原之一。

英文摘要:

Objective To develop arapid, 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-o ne 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 immunofluo rescent assay.Results Only the cDNA from HRV was po sitive 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 po sitive rates, it was found53.3% in phary ngitis patients;43.8% in laryngitis patients and 28.7 % in bronchitis pa tients.In Sep.2002 and from Aug.2003 to Oct.2003, HRV positive rates w ere high(21.6%-32.6 %), w ith Sep.2003 in particular — 32.6 %.From Mar.2003 to Jul.2003, HRV positive rates maintained from 16.0% to19.1%.Conclusion　HRV was one of the impo rtant agents for acute respiratory infections in infants and young children in Beijing.

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