摘要
目的 从北京患急性呼吸道感染患儿的临床标本中分离人偏肺病毒（HMPV）,推动对该病毒的深入研究.方法 2008年5月至2009年4月收集门诊和病房因急性呼吸道感染患儿的标本,经逆转录-聚合酶链反应（RT-PCR）检测核酸和直接免疫荧光法检测抗原筛选到HMPV阳性标本后接种传代的恒河猴肾细胞（LLC-MK2）,经37 ℃和33 ℃孵育后用直接免疫荧光法检测细胞内的HMPV抗原,得到阳性结果后经RT-PCR进一步确定基因型.结果 从1092例病房和门诊收集的急性呼吸道患儿标本中用RT-PCR技术检测到81例为HMPV阳性,总阳性率为7.4%（81/1092）.取其中33例接种到LLC-MK2细胞进行HMPV分离,有5份标本的接种细胞中可检测到HMPV抗原,并经RT-PCR检测进一步证实,分离株与常见的呼吸道病毒抗体无交叉反应.24 h内新鲜接种的标本更易分离到病毒.分离到的毒株有4株为A基因型,1株为B基因型.结论 从北京急性呼吸道感染患儿的标本中分离到两种基因型的HMPV,将进一步推动对该病毒的研究.
Objective To obtain isolated human metapneumovirus （HMPV） strains from clinical specimens collected from infants and children in Beijing and to promote the investigation on this important respiratory pathogen. Method Clinical specimens including throat swabs from outpatients and nasopharyngeal aspirates from hospitalized children were collected from infants and children visited the affiliated children＇s hospital for acute respiratory infections during May 2008 to April 2009. HMPV positive specimens identified by RT-PCR and/or direct immunofluorescent assay with monoclonal antibody against HMPV were inoculated to LLC-MK2 cells and incubated at 37 ℃ and 33 ℃, respectively. The replication of the virus in the cells was detected by direct immunofluorescent assay followed by RT-PC. The genotypes of the isolated virus strains were identified by RT-PCR. Result Out of 1092 clinical specimens, 81 were HMPV positive by RT-PCR, the positive rate was 7.4% （81/1092）. Among these positive specimens, 33 were inoculated to LLC-MK2 cells and the replication of HMPV was revealed by antigen detection and RTPCR from 5 out of these 33 inoculates. These isolated viruses could be passed in LLC-MK2 cells and were not cross-reacted with other common respiratory viruses, such as ADV, RSV and Parainfluenza viruses 1/2/3by monoclonal antibodies against these viruses in direct immunofluorescent assay. The HMPV was more likely to be isolated from fresh specimens within 24 hours after the collection of specimens which were not frozen. Four of the 5 isolated strains were identified as genotype A and 1 as genotype B. Unlike other respiratory viruses, these isolated HMPV did not show specific CPE in cell culture and the replication of the virus was identified by antigen detection and RT-PCR. Conclusion HMPV of both genotypes were isolated from infants and children with acute respiratory infections in Beijing which will accelerate the investigation of this important virus.
出处
《中华儿科杂志》
CAS
CSCD
北大核心
2010年第11期820-823,共4页
Chinese Journal of Pediatrics
基金
国家自然科学基金（30872153）
关键词
儿童
呼吸道感染
人偏肺病毒
Child
Respiratory tract infections
Human metapneumovirus
