建立检测18种(型/亚型)呼吸道病毒的两种循环参数的实时荧光定量PCR

被引：7

作者：

陈钰静

丁细霞

郭勇晖

余楠

潘玉先

余志武

陈满君

王压娣

杜晓棠

车小燕

机构：

[1] 南方医科大学珠江医院检验医学部

来源：

临床检验杂志 | 2014年 / 32卷 / 05期

关键词：

呼吸道病毒; 实时荧光定量聚合酶链反应; 特异性; 敏感性;

D O I：

10.13602/j.cnki.jcls.2014.05.06

中图分类号：

R450 [];

学科分类号：

100215 ;

摘要：

目的建立两种不同循环参数的实时荧光定量PCR,检测包括流感病毒、冠状病毒等18种（型/亚型）呼吸道病毒。方法引用文献报道的18种（型/亚型）呼吸道病毒引物和探针,建立两种不同循环参数的实时荧光定量PCR法,考核方法的敏感性、特异性和准确性,并用临床标本进行验证。结果成功建立了可检测18种（型/亚型）呼吸道病毒的两种不同循环参数的实时荧光定量PCR法;该法特异性较好,最低检测限为10拷贝数/μL,变异系数（CV）为1.26%3.43%,重复性良好;408份临床标本中,用该法检测鼻拭子标本阳性率为63.2%（225/356）,肺泡灌洗液标本阳性率为25.0%（13/52）。结论建立的实时荧光定量PCR法简便、特异、敏感、稳定,适用于临床常见呼吸道病毒感染的早期诊断。

引用

页码：340 / 344

页数：5

共 12 条

[1]

Real-time PCR assays for detection of bocavirus in human specimens. Lu Xiaoyan,Chittaganpitch Malinee,Olsen Sonja J,Mackay Ian M,Sloots Theo P,Fry Alicia M,Erdman Dean D. Journal of Clinical Microbiology . 2006

[2]

Increased detection of respiratory syncytial virus, influenza viruses, parainfluenza viruses, and adenoviruses with real-time PCR in samples from patients with respiratory symptoms. van de Pol Alma C,van Loon Anton M,Wolfs Tom F W,Jansen Nicolaas J G,Nijhuis Monique,Breteler Els Klein,Schuurman Rob,Rossen John W A. Journal of Clinical Microbiology . 2007

[3]

A One-Step, Real-Time PCR Assay for Rapid Detection of Rhinovirus[J] . Duc H. Do,Stella Laus,Amy Leber,Mario J. Marcon,Jeanne A. Jordan,Judith M. Martin,Robert M. Wadowsky. &nbspThe Journal of Molecular Diagnostics . 2010 (1)

[4]

One-step real-time reverse transcription-PCR assays for detecting and subtyping pandemic influenza A/H1N1 2009, seasonal influenza A/H1N1, and seasonal influenza A/H3N2 viruses[J] . Mina Nakauchi,Yoshihiro Yasui,Tatsuya Miyoshi,Hiroko Minagawa,Tomoyuki Tanaka,Masato Tashiro,Tsutomu Kageyama. &nbspJournal of Virological Methods . 2010 (1)

[5] Impact of human coronavirus infections in otherwise healthy children who attended an emergency department [J].

Esposito, Susanna ;

Bosis, Samantha ;

Niesters, Hubert G. M. ;

Tremolati, Elena ;

Begliatti, Enrica ;

Rognoni, Alessandro ;

Tagliabue, Claudia ;

Principi, Nicola ;

Osterhaus, Albert D. M. E. .

JOURNAL OF MEDICAL VIROLOGY, 2006, 78 (12) :1609-1615

[6]

Design and performance testing of quantitative real time PCR assays for influenza A and B viral load measurement[J] . C.L Ward,M.H Dempsey,C.J.A Ring,R.E Kempson,L Zhang,D Gor,B.W Snowden,M Tisdale. &nbspJournal of Clinical Virology . 2003 (3)

[7]

Applicability of a real-time quantitative PCR assay for diagnosis of respiratory syncytial virus infection in immunocompromised adults. van Elden L J R,van Loon A M,van der Beek A,Hendriksen K A W,Hoepelman A I M,van Kraaij M G J,Schipper P,Nijhuis M. Journal of Clinical Microbiology . 2003

[8] Rapid and quantitative detection of human adenovirus DNA by real-time PCR [J].

Heim, A ;

Ebnet, C ;

Harste, G ;

Pring-Åkerblom, P .

JOURNAL OF MEDICAL VIROLOGY, 2003, 70 (02) :228-239

[9]

Frequent detection of human coronaviruses in clinical specimens from patients with respiratory tract infection by use of a novel real-time reverse-transcriptase polymerase chain reaction. van Elden Leontine J R,van Loon Anton M,van Alphen Floris,Hendriksen Karin A W,Hoepelman Andy I M,van Kraaij Marian G J,Oosterheert Jan-Jelrik,Schipper Pauline,Schuurman Rob,Nijhuis Monique. The Journal of Infectious Diseases . 2004

[10]

A previously undescribed coronavirus associated with respiratory disease in humans. Ron A. M. Fouchier,Nico G. Hartwig,Theo M. Bestebroer,Berend Niemeyer,Jan C. de Jong,James H. Simon,Albert D. M. E. Osterhaus. Proceedings of the National Academy of Sciences of the United States of America . 2004

← 1 2 →