荧光RT-PCR对乙型流感病毒爆发的检测

Detection of influenza B virus Victoria lineages outbreaks by real-time PCR

目的探讨荧光RT-PCR方法对乙型流感病毒爆发的诊断价值。方法采集暴发性疫情的流感样病例的咽拭子,提取病毒RNA,PCR方法分别检测甲型和乙型流感病毒的核酸。然后,对核酸检测阳性标本进行MDCK培养,经红细胞凝集和凝集抑制试验对病毒进行鉴定和分型。结果 18份咽拭子中15份乙型流感病毒核酸呈阳性,甲型流感病毒未检出;15份乙型流感病毒核酸阳性标本经MDCK培养,9份标本发生细胞病变,经鉴定和分型,都属于乙型流感病毒Victoria系。结论 RT-PCR方法对乙型流感病毒爆发的检测具有快速、敏感和高度特异性的特点。本地区爆发的乙型流感病毒为乙型流感病毒Victoria系,提示在今后决定我国疫苗成份时应考虑地区流行的特点。

Objective To evaluate the real-time PCR assays for accurate diagnosis of influenza B virus outbreaks.Methods The swallow swabs from the influenzalike cases of suspected influenza outbreaks in five primary school of Qingdao city were collected.The viral RNA was extracted（QIAGEN,viral RNA kit） to amplify the specific regions of influenza A and B virues by RT-PCR,respectively.The virus of PCR positive samples were isolated by MDCK（Madin-Darby canine kidney cell）,and then identified by hemagglutination inhibition test.Results None of specimens was influenza A positive.But 15 of 18 swallow swabs were influenza B positive.And all of 9 stains isolated from 15 positive samples by MDCK method were identified to be influenza virus subtype B Victoria lineages by hemagglutination inhibition test.Conclusion The real-time PCR assay was a quick,sensitive and specific method for molecular diagnosis of influenza B virus.The assay was successfully applied to 18 primary samples collected in Qingdao city.This influenza outbreak demonstrates the need for national or regional control authorities approve the composition and formulation of vaccines according to recent epidemiological evidence in each district.