中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2010年
4期
204-208
,共5页
王长兵%游爱萍%肖密丝%朱冰
王長兵%遊愛萍%肖密絲%硃冰
왕장병%유애평%초밀사%주빙
流感病毒,A型%核壳蛋白质类%酶联免疫吸附测定
流感病毒,A型%覈殼蛋白質類%酶聯免疫吸附測定
류감병독,A형%핵각단백질류%매련면역흡부측정
Influenza A virus%Nuclocapsid proteins%Enzyme-linked immunosorbent assay
目的 研制检测靶位针对甲型流行性感冒(流感)病毒核蛋白的甲型流感病毒抗原检测试剂盒,建立能检测甲型流感病毒所有亚型的方法.方法 采用双抗体夹心法检测标本中的甲型流感病毒,建立甲型流感病毒抗原ELISA方法,并对该试剂盒的灵敏性、特异性、精确性、稳定性进行测试和临床考核.结果 甲型流感病毒抗原ELISA试剂盒核蛋白检测下限为7.63 ng/Ml,灵敏度是血球凝集方法的256倍,是美国Quickdel公司胶体金产品的16倍;与乙型流感病毒、呼吸道合胞病毒、呼吸道腺病毒、副流感病毒Ⅰ/Ⅲ型、肺炎支原体、鸡新城疫病毒、鸡法氏囊病毒、鸡传染性支气管炎病毒均无交叉,特异度为100%;批内、批间变异系数(CV)值均<15%,符合国家标准;在4℃以下的稳定性可达1年,能通过37℃7 d加速试验.可检测亚型分别为H1N1、H3N2、H5N1和H9N2的甲型流感病毒.人流感病毒临床试验表明,与常规细胞培养方法比较,阳性符合率为93.44%,阴性符合率为99.31%;禽流感病毒临床试验表明,与常规细胞培养方法比较,阳性符合率为95.45%,阴性符合率为98.09%.结论 甲型流感病毒抗原ELISA试剂盒灵敏性、特异性强,可用于人甲型流感病毒和禽流感病毒感染的流行病学调查.
目的 研製檢測靶位針對甲型流行性感冒(流感)病毒覈蛋白的甲型流感病毒抗原檢測試劑盒,建立能檢測甲型流感病毒所有亞型的方法.方法 採用雙抗體夾心法檢測標本中的甲型流感病毒,建立甲型流感病毒抗原ELISA方法,併對該試劑盒的靈敏性、特異性、精確性、穩定性進行測試和臨床攷覈.結果 甲型流感病毒抗原ELISA試劑盒覈蛋白檢測下限為7.63 ng/Ml,靈敏度是血毬凝集方法的256倍,是美國Quickdel公司膠體金產品的16倍;與乙型流感病毒、呼吸道閤胞病毒、呼吸道腺病毒、副流感病毒Ⅰ/Ⅲ型、肺炎支原體、鷄新城疫病毒、鷄法氏囊病毒、鷄傳染性支氣管炎病毒均無交扠,特異度為100%;批內、批間變異繫數(CV)值均<15%,符閤國傢標準;在4℃以下的穩定性可達1年,能通過37℃7 d加速試驗.可檢測亞型分彆為H1N1、H3N2、H5N1和H9N2的甲型流感病毒.人流感病毒臨床試驗錶明,與常規細胞培養方法比較,暘性符閤率為93.44%,陰性符閤率為99.31%;禽流感病毒臨床試驗錶明,與常規細胞培養方法比較,暘性符閤率為95.45%,陰性符閤率為98.09%.結論 甲型流感病毒抗原ELISA試劑盒靈敏性、特異性彊,可用于人甲型流感病毒和禽流感病毒感染的流行病學調查.
목적 연제검측파위침대갑형류행성감모(류감)병독핵단백적갑형류감병독항원검측시제합,건립능검측갑형류감병독소유아형적방법.방법 채용쌍항체협심법검측표본중적갑형류감병독,건립갑형류감병독항원ELISA방법,병대해시제합적령민성、특이성、정학성、은정성진행측시화림상고핵.결과 갑형류감병독항원ELISA시제합핵단백검측하한위7.63 ng/Ml,령민도시혈구응집방법적256배,시미국Quickdel공사효체금산품적16배;여을형류감병독、호흡도합포병독、호흡도선병독、부류감병독Ⅰ/Ⅲ형、폐염지원체、계신성역병독、계법씨낭병독、계전염성지기관염병독균무교차,특이도위100%;비내、비간변이계수(CV)치균<15%,부합국가표준;재4℃이하적은정성가체1년,능통과37℃7 d가속시험.가검측아형분별위H1N1、H3N2、H5N1화H9N2적갑형류감병독.인류감병독림상시험표명,여상규세포배양방법비교,양성부합솔위93.44%,음성부합솔위99.31%;금류감병독림상시험표명,여상규세포배양방법비교,양성부합솔위95.45%,음성부합솔위98.09%.결론 갑형류감병독항원ELISA시제합령민성、특이성강,가용우인갑형류감병독화금류감병독감염적류행병학조사.
Objective To develop and verify an influenza A virus antigen-detecting kit which can detect all the subtypes of influenza A virus. Methods Double-antibodies sandwich enzyme linked immunosorbent assay (ELISA) was utilized for developing the influenza A virus antigen-detecting kit. The sensitivity, specificity, accuracy and stability of the kit were evaluated by the clinical samples. Results The lower detection limit of the kit for the N protein was 7. 63 ng/mL, which was 256 times lower than that of the hemagglutination and 16 times lower than that of immune colloidal gold technique. The kit didn't show any cross-reaction with the influenza B virus, respiratory syncytial virus, respiratory adenovirus, para-influenza virus type Ⅰ and Ⅲ, mycoplasma pneumomiae, avian newcastle disease virus, avian infectious bursal disease virus or avian infectious bronchitis virus. The specificity was 100%. Both the intra batch variaton coefficient (CV) value and inter batch CV value were less than 15%, which met the national standard for ELISA kits. The results proved that the kit could keep stable at 4 ℃ for more than 1 year and at 37 ℃ for more than 7 days. The kit could identify H1N1, H3N2, H5N1 and H9N2 influenza A viruses. The clinical research data of human influenza virus showed the consistency rate between the kit and regular cell culture method was 93. 44% for the positive samples and 99. 31% for the negative samples. The clinical research data of avian influenzavirus showed the consistency rate between the kit and regular cell culture method was 95. 45% for positive samples and 98. 09% for negative samples. Conclusion The influenza A virus antigen-detecting ELISA kit can be used for the epidemiological survey of the infection of human influenza A virus or avian influenza virus with high sensitivity and specificity.
