Detection of antibody to avian influenza A (H5N1) virus in human serum by using a combination of serologic assays
Detection of antibody to avian influenza A (H5N1) virus in human serum by using a combination of serologic assays
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DOI：
10.1128/jcm.37.4.937-943.1999
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发表时间：
1999-04-01
影响因子：
9.4
通讯作者：
Katz, JM
中科院分区：
文献类型：
--
作者：
Rowe, T;Abernathy, RA;Katz, JM
在1997年5月至12月期间，香港共发现18宗由甲型H5N1禽流感病毒引起的轻微至严重呼吸系统疾病。禽流感病毒在人类中的出现促使进行流行病学调查，以确定病毒在人与人之间传播的程度以及与感染有关的风险因素。血凝抑制试验是检测人类流感病毒感染的血清学标准方法，但对禽流感病毒诱导的抗体的检测敏感性较低。因此，我们发展了一种更灵敏的微量中和试验来检测人类禽流感抗体。血凝抑制试验和微量中和试验的直接比较表明，后者在检测受感染个体中的人类H5N1病毒抗体方面要敏感得多。还建立了MS特异性间接酶联免疫吸附试验(EL ISA)检测儿童血清。比较了微量中和试验与MS特异性间接酶联免疫吸附试验的敏感性和特异性。当与确证的MS特异性Western印迹试验相结合时，两种检测方法的特异性都得到了提高。用微量中和试验结合免疫印迹法检测18~59岁成人抗H5抗体的灵敏度(80%)和特异度(96%)最高。用该方法检测15岁以下儿童血清中抗H5抗体的灵敏度和特异度分别为100%和100%。这一新的测试算法正用于禽流感H5N1暴发的血清流行病学调查。
From May to December 1997, 18 cases of mild to severe respiratory illness caused by avian influenza A (H5N1) viruses were identified in Hong Kong. The emergence of an avian virus in the human population prompted an epidemiological investigation to determine the extent of human-to-human transmission of the virus and risk factors associated with infection. The hemagglutination inhibition (HI) assay, the standard method for serologic detection of influenza virus infection in humans, has been shown to be less sensitive for the detection of antibodies induced by avian influenza viruses. Therefore, we developed a more sensitive microneutralization assay to detect antibodies to avian influenza in humans. Direct comparison of an HI assay and the microneutralization assay demonstrated that the latter was substantially more sensitive in detecting human antibodies to H5N1 virus in infected individuals. An MS-specific indirect enzyme-linked immunosorbent assay (ELISA) was also established to test children's sera. The sensitivity and specificity of the microneutralization assay were compared with those of an MS-specific indirect ELISA. When combined with a confirmatory MS-specific Western blot test, the specificities of both assays were improved. Maximum sensitivity (80%) and specificity (96%) for the detection of anti-H5 antibody in adults aged 18 to 59 years were achieved by using the microneutralization assay combined with Western blotting. Maximum sensitivity (100%) and specificity (100%) in detecting anti-H5 antibody in sera obtained from children less than 15 years of age were achieved by using ELISA combined with Western blotting. This new test algorithm is being used for the seroepidemiologic investigations of the avian H5N1 influenza outbreak.