中国2005年脊髓灰质炎实验室网络的运转与监测

Surveillance and Running Status of Laboratories Network for Poliomyelitis in 2004 in China.

目的利用中国(未包括香港、澳门特别行政区和台湾地区)2005年维持无脊髓灰质炎(脊灰)状态的实验室资料,评估脊灰实验室网络的运转与监测情况。方法分析31个省(自治区、直辖市,下同)急性弛缓性麻痹(AFP)病例监测系统经计算机联网上报的AFP病例个案调查表数据库和中国疾病预防控制中心(CDC)病毒病预防控制所国家脊灰实验室监测数据库。结果2005年31个省CDC从5 195例AFP病例中收集了10 340份粪便标本,所有粪便标本在L20B、RD细胞上同时进行病毒分离。从293例AFP病例粪便标本中分离到脊灰病毒(PV),分离率为5.64%;从551例AFP病例粪便标本中分离到非脊灰肠道病毒(NPEV),分离率为10.61%。国家脊灰实验室共收到各省CDC脊灰实验室送检的404株PV标本;其中293株分离于AFP病例,其余分离于AFP病例接触者、流动人口和健康人群。对404株PV使用聚合酶链反应-限制性酶切片段长度多态性分析(PCR-RFLP)进行型内鉴定,PVⅠ型疫苗株72株,PVⅡ型疫苗株138株,PVⅢ型疫苗株74株,混合型疫苗株91株,PV+NPEV 12株,Ⅰ、Ⅱ、Ⅲ型疫苗变异株分别为6、6、5株。将PV混合株进行单型分离后,进行酶联免疫吸附试验(ELISA)型内鉴定,共鉴定单型毒株521株,其中疫苗类似株(SL)498株,占所有送检毒株的95.59%;非疫苗类似株(NSL)2株,占0.38%;双反应株(DRV)18株,占3.45%;阴性2株;无效1株。所有型内鉴定异常和部分从高危零剂次服苗者分离的118株,做了VP1编码区全基因的序列测定和分析。在江苏、安徽省CDC陆续送检的编号为9230病例的33份病毒分离物(其中江苏省2份,安徽省31份),连续8个月从安徽省舒城县1例免疫缺陷患儿分离到疫苗衍生脊灰病毒(iVDPV);VP1编码区全基因的序列测定图谱显示,19株Ⅱ型iVDPVs,13株Ⅲ型iVDPVs。2005年世界卫生组织(WHO)使用5份盲样标本进行病毒分离职能考核,31个省CDC脊灰实验室成绩均为满分。对14个省CDC脊灰实验室进行了现场考核,全部通过认证。结论中国2005年继续维持无脊灰状态,脊灰实验室网络运转正常,为维持无脊灰提供了完整的实验室依据。

Objective In order to evaluate the running status and maintain the laboratories network for poliomyelitis in China, and provide the laboratory data for ＂Polio free＂ in China. The surveillance and running status of laboratories network for poliomyelitis in China in 2005 was analyzed. Methods The Acute flaccid paralysis （AFP） cases investigation database reported by AFP surveillance system of 31 provinces and national laboratory database were used. Results 10,340 stool samples collected from 5195 AFP cases by 31 provincial CDC, and all stool samples were made viral isolation on both L20B cell and RD cell according to WHO manual. 293 polioviruses were isolated from AFP cases, the isolation rate is 5.6% ; and 551 of non-polio enteroviruses （NPEV） were isolated from AFP cases,the isolation rate is 10.7 %. We retest the serotype of all the polio positive isolates sent by 31 provincial laboratories and done PCR-RFLP intratypic identification tests. Among 404 isolates, 72 of type 1 vaccine-like viruses, 138 of type 2 vaccine-like viruses, 74 of type 3 vaccine-like viruses, 91 of mixed viruses, 12 of vaeein-like viruses mixed with enteroviruses, 17 viruses were identified as mutant viruses. We also do another intratypic identification tests,ELISA test, according to WHO manual, among total 521 single serotype viruses, 498 of sabin-like viruses（SL）, 2 of non-sabin-like viruses （NSL）, 18 of double reaction viruses（DRV）, all results with mutant viruses of PCR test, results with DRV and NSL of ELISA test and part of the polioviruses isolated from patients with 0 dose immunization were selected for VP1 sequencing, total 118 viruses were done VPI sequencing, 19 of type 2 iVDPVs and 13 type 3 iVDPVs were found in, Anhui Province. 5 blind specimens for proficiency test was distributed by World Health Organization （WHO）, all provincial laboratories for poliomyelitis got full scores and passed the proficiency test. 14 provincial laboratories for poliomyelitis also got accreditation by experts through on-site review. Conclusion ＂Polio-free＂ status were maintained in 2005 in China, the laboratories for poliomyelitis network ran smoothly, the laboratory surveillance data was detailed and veracious, the surveillance system is sensitive enough, it can provide complete laboratory material for maintaining the polio-free in China.