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

Abstract

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].Chinese journal of Epidemiology,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

Received:April 28, 2005

DOI：

KeyWord: 鼻病毒巢式逆转录聚合酶链反应

English Key Word: Human rhinovirus Nested reverse transcription polymerase chain reaction

FundProject: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)

Author Name	Affiliation	E-mail
Zhao Linqing	Laboratory of Virology, Beijing Municipal Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China
Qian Yuan	Laboratory of Virology, Beijing Municipal Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China	yqianbjc@263.net
Zhu Runan	Laboratory of Virology, Beijing Municipal Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China
Deng Jie	Laboratory of Virology, Beijing Municipal Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China
Wang Fang	Laboratory of Virology, Beijing Municipal Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China

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Abstract:

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。

English Abstract:

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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