流感病毒Real—time RT—PCR核酸檢測及流行情況分析

摘要：目的 分析鄂爾多斯市2012年流感檢測結(jié)果，

關(guān)鍵詞：流感監(jiān)測；病毒核酸；Real-time RT-PCR

Detection of Influenza Viruses Using Real-time RT-PCR and Analysis of Influenza Epidemic Situation

JI Lin-li

（Center For Disease Control and Prevention of Erdos， Erdos 017000，Inner Mongolia，China）

Abstract：Objective To analyze the results of detection on influenza virus in Ordos in 2012， in order to provide scientific basis for prevention and control of flu in this area. Methods Influenza throat swab specimens were collected from sentinel hospital for virus nucleic acid detection by Real-time RT-PCR. Results： A total of 281 throat swab specimens were detected， 35 were positive for influenza virus nucleic acid， at a percentage of 12.46%. In which there was 16 cases of B type flu， at a percentage of 5.69%， 15 cases of seasonal H3， at a percentage of 5.34%，2 cases of new H1N1， at a percentage of 0.71%， 2 positive of A type undentified，at a percentage of 0.71%. The detection positive rate were 11.36%、27.50%、37.50% in January ，F(xiàn)ebruary and March， were 5.00% in October and December，no positive sample was detected in other month. Conclusion The survey showed that the major epidemic seasons were winter and spring，the major epidemic strains of the flu were B type and H3 type.

Key words：Surveillance of influenza ； Virus nucleic acid； Real-time RT-PCR

流行性感冒簡稱流感，是由流感病毒引起的急性上呼吸道傳染病。流感潛伏期短，傳播迅速，且抗原易變異，人群對變異株普遍易感，因此控制難度大[1]。目前，對人類危害較重的流感病毒株是A型和B型，其中A型流感病毒抗原變異率高，容易造成世界范圍內(nèi)大流行[2]。2009年，甲型H1N1流感就是由于流感病毒變異形成的一種與豬流感病毒高度同源的新型變異株引起的流感大流行[3]。通過流行病學(xué)和病原學(xué)監(jiān)測，可以幫助做好流感流行趨勢和優(yōu)勢毒株的預(yù)測，及早采取相應(yīng)措施，對防止流感流行具有重要意義[4]。鄂爾多斯市疾病預(yù)防控制中心流感監(jiān)測網(wǎng)絡(luò)實(shí)驗室2012年全年共對281份流感病例咽拭子標(biāo)本進(jìn)行了病毒核酸的檢測，現(xiàn)將結(jié)果報告如下：

1資料與方法

1.1一般資料

1.1.1標(biāo)本來源 哨點(diǎn)醫(yī)院（鄂爾多斯市中心醫(yī)院）按照《全國流感監(jiān)測方案（2010版）》要求采集流感樣病例咽拭子標(biāo)本。流感樣病例指發(fā)熱（體溫≥38℃），伴有咳嗽或咽疼癥狀之一，同時缺乏其他的實(shí)驗室診斷依據(jù)[5]。

1.1.2主要試劑和儀器 流感病毒RNA提取試劑盒（QIAamp viral RNA Mini Kit ）為德國QIAGEN公司產(chǎn)品、Real-time RT-PCR擴(kuò)增試劑盒（AM1005）為美國 ABI公司產(chǎn)品、檢測引物由上海生工合成；主要檢測儀器有ABI Step-one plus型熒光擴(kuò)增儀、QL-866型震蕩器，等。

1.2方法

1.2.1核酸提取 按照QIAGENR Neasy Mini kit 操作說明進(jìn)行。

1.2.2 Real-time RT-PCR檢測 Real-time RT-PCR反應(yīng)體系配置按AgPath-IDTM One-Step RT-PCR Kit說明進(jìn)行，反應(yīng)程序參照《甲型H1N1流感監(jiān)測方案（第二版）》[5]。

1.2.3結(jié)果判定 在陰陽對照成立下，CT值≤37且有典型的S型曲線報告為檢測結(jié)果陽性。……