| 龚成,罗明,陈萌,张铁钢,张合润,王玉梅,李仁清,董梅,陈维新,陈丽娟.Ⅰ型脊髓灰质炎野病毒实时荧光RT-PCR快速检测方法的评价[J].Chinese journal of Epidemiology,2012,33(7):726-729 |
| Ⅰ型脊髓灰质炎野病毒实时荧光RT-PCR快速检测方法的评价 |
| Study on the Fast Testing Strategy for identifying the wild poliovirus Ⅰ |
| Received:December 26, 2011 |
| DOI： |
| KeyWord: 脊髓灰质炎病毒 脊髓灰质炎野病毒 实时荧光RT-PCR |
| English Key Word: Poliovirus Poliomyelitis, wild poliovirus Real time RT-PCR |
| FundProject: |
| Author Name | Affiliation | E-mail | | GONG Cheng | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | LUO Ming | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | CHEN Meng | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | ZHANG Tie-gang | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | ZHANG He-run | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | WANG Yu-mei | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | LI Ren-qing | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | DONG Mei | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | CHEN Wei-xin | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | | | CHEN Li-juan | The Immunization Institute, Beijing Center for Disease Control and Prevention, Beijing 100013, China | bjcdccblj@hotmail.com |
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| Abstract: |
| 目的 评价和探讨Ⅰ型脊髓灰质炎(脊灰)野病毒快速检测策略.方法 采集671名来自脊灰野病毒流行疫区的在京学生粪便标本,采用一组实时荧光RT-PCR代替标准方法分别进行肠道病毒通用型检测、脊灰病毒分型检测、Ⅰ型脊灰疫苗株和野毒株鉴别检测.并利用Ⅰ、Ⅱ和Ⅲ型脊灰病毒标准株(Sabin株)和Ⅰ型野毒株对实时荧光RT-PCR和标准方法的灵敏度进行比较和评价.结果 (1)新脊灰病毒检测流程(肠道病毒通用型检测+脊灰病毒分型检测+脊灰野毒鉴别检测)检出非脊灰肠道病毒33例；脊灰病毒疫苗株16例,其中Ⅰ型5例、Ⅱ型1例、Ⅲ型3例、Ⅰ+Ⅱ型1例、Ⅰ+Ⅲ型4例、Ⅰ+Ⅱ+Ⅲ型2例；脊灰病毒Ⅰ型野毒株3例.(2)3种实时荧光RT-PCR检测脊灰病毒比标准方法敏感1～ 100倍.其中肠道通用型检测和脊灰病毒分型检测2种方法对Ⅱ型脊灰病毒疫苗株的检测,比标准方法敏感100倍；脊灰病毒分型检测能准确区分Ⅰ、Ⅱ和Ⅲ型脊灰疫苗株,脊灰野毒株鉴别检测对Ⅰ型疫苗株的检测比标准方法敏感10倍；3种实时荧光RT-PCR均能有效检测出Ⅰ型脊灰野毒株,比标准方法敏感10倍.结论 新的脊灰病毒检测流程,可大幅缩短检测时间,提高检测通量,且灵敏度高于标准方法,适合于脊灰应急检测应对突发疫情. |
| English Abstract: |
| Objective To explore the Fast Testing Sstrategy (FTS) for wild poliovirus Ⅰ (WP1).Methods Epidemiological investigations were carried out on 671 students from WP1 epidemic areas in China.A set of real time RT-PCR assays,including panenterovirus testings (PE) assay,poliovirus serotypings(PS) assay and the assay distinguishing wild strain from vaccine strain of poliovirus Ⅰ (DWV) were introduced into the screening program for WPV1 to replace the conventional RT-PCR,recommended by the China National Polio Laboratory (GNPL).Additionally,sensitivities of all the assays were assessed by poliovirus type Ⅰ to Ⅲ (Sabin stain) and the isolated WPV I.Results ( 1 ) 33 non-poliovirus enterovirus (NPEV) cases were detected,with 16 polio vaccinerelated cases including 5 polio Ⅰ,1 polio Ⅱ,3 polio Ⅲ,1 polio Ⅰ +Ⅱ,4 polio Ⅰ + Ⅲ and 2 polio Ⅰ + Ⅱ + Ⅲ.Three WPV 1 cases were also detected in this study and confirmed by GNPL.(2) For polio virus vaccine strain,sensitivities of the set of real time RT-PCR assays ranged from 1 to 100 times than that of the in-housc RT-PCR assay.The sensitivities of PE and PS assays for the detection of polio Ⅱ were 100 times than that of the RT-PCR assay and the sensitivity of DWV assay used for the detection of polio Ⅰ were 10 times than that of the RT-PCR assay.For WPV1,the sensitivity of three real time RT-PCR was 10 times higher than that of the RT-PCR assay.Conclusion The novel FTS for WPV I suggested by this study would include PE,PS and DWV.It not only could greatly shorten the testing time but also more sensitive than the RT-PCR and suited for emergency detection for WPV1. |
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