离心培养直接荧光染色法在流感病毒检测中的应用

Monitoring of influenza by shell viral assay and direct immunofluorescent staining

目的 对北京地区 5家医院的流感情况进行前瞻性调查 ,采用离心培养技术快速检测流感病毒 ,用于流感的诊断和分型。方法 分别收集北京地区 2 0 0 0～ 2 0 0 1年及 2 0 0 2年具有流感样症状患者的鼻吸液标本 713份和 89份 ,离心后接种于单层犬肾细胞 (MDCK)的 2 4孔板内 ,采用荧光单抗技术鉴定培养物甲、乙分型。结果 2 0 0 0～ 2 0 0 1年甲、乙型流感病毒培养阳性率分别为 2 6 2 %和 15 3 % ,2 0 0 2年甲、乙型流感病毒培养阳性率分别为 2 2 %和 11 2 %。结论 离心培养及荧光单抗染色技术能够快速确诊流感患者 ,及时提供流感病毒分型报告。

Objective To establish a rapid culture method for monitoring of influenza circulation and laboratory diagnosis of individual patients with influenza. Methods Nasal aspirate specimens were spun onto 24-well plate containing confluent monolayers of Madin-Darby Canine Kidney (MDCK) cells. Fluorescein labeled monoclonal antibodies to influenza A and B viruses were used to stain and type the isolates. Results A total of 713 nasal aspirates specimens from patients experiencing influenza-like symptoms was collected in Beijing between December of 2000 and March of 2001, 187(26 2%) specimens were found with influenza A virus and 109(15 3%) with influenza B virus. 2 2% with influenza A and 11 2% with influenza B virus were detected in 89 nasal aspirates specimens between Jan-Apr 2002. Conclusion Prevalence of influenza was low during 2000～2002 year non-epidemic period in Beijing. The combination of shell viral assay and direct immunofluorescent staining can provide a rapid laboratory diagnosis of influenza, which makes possible for the patients to receive treatment of anti-influenza virus drugs.