Daum Luke T, Canas Linda C, Schadler Cassandra A, Ujimori Veronica A H, Huff William B, Barnes William J, Lohman Kenton L
Molecular Epidemiology Branch, 2601 West Gate Road, Brooks AFB, San Antonio, TX 78235-5241, USA.
J Clin Virol. 2002 Dec;25(3):345-50. doi: 10.1016/s1386-6532(02)00043-4.
Influenza is a viral respiratory pathogen responsible for frequent seasonal epidemics. There are currently three major human influenza viruses in global circulation, H1N1, H3N2 and B.
A one-step multiplex reverse transcription (RT)-polymerase chain reaction (PCR) assay targeting the HA1 segment of the human hemagglutinin gene was developed as a rapid surveillance method.
A researcher-blind study was performed using 112 randomly selected, culture-positive clinical samples collected through the Department of Defense (Global Emerging Infectious Surveillance (DOD-GEIS) influenza network during the 2000-2001 influenza season. Three subtype specific primer sets capable of producing PCR products with base-pair lengths of 585, 402 and 290 corresponding to influenza H1, H3, and B subtypes, respectively, were utilized together in a one step, one tube, reaction.
Multiplex primers were able to simultaneously type, and subtype 100% (112/112) of positive cultures.
The results confirm that this assay is a highly sensitive and timely surveillance tool for rapid detection and simultaneous subtyping of clinical influenza specimens isolated worldwide.
流感是一种病毒性呼吸道病原体，可引发频繁的季节性流行。目前全球流行的主要人类流感病毒有三种，即H1N1、H3N2和B型。
开发一种针对人血凝素基因HA1片段的一步多重逆转录（RT）-聚合酶链反应（PCR）检测方法，作为一种快速监测方法。
在2000 - 2001年流感季节，使用通过国防部（全球新兴传染病监测（DOD-GEIS）流感网络）随机选取的112份培养阳性临床样本进行了一项研究者盲法研究。三种亚型特异性引物组能够分别产生对应流感H1、H3和B亚型的碱基对长度为585、402和290的PCR产物，将它们在一步、一管反应中共同使用。
多重引物能够同时对100%（112/112）的阳性培养物进行分型和亚型鉴定。
结果证实该检测方法是一种高度灵敏且及时的监测工具，可用于快速检测和同时对全球分离的临床流感样本进行亚型鉴定。
