以哈维氏弧菌GYC1108-1细菌颗粒性抗原免疫8周龄雌性BALB/c小鼠,5次免疫后取其脾细胞与SP2/0-Ag-14骨髓瘤细胞融合。用间接酶联免疫吸附试验(ELISA)对杂交瘤进行筛选,阳性克隆经3次亚克隆后,获得一株针对GYC1108-1脂多糖(LPS)的特异性...以哈维氏弧菌GYC1108-1细菌颗粒性抗原免疫8周龄雌性BALB/c小鼠,5次免疫后取其脾细胞与SP2/0-Ag-14骨髓瘤细胞融合。用间接酶联免疫吸附试验(ELISA)对杂交瘤进行筛选,阳性克隆经3次亚克隆后,获得一株针对GYC1108-1脂多糖(LPS)的特异性单克隆抗体,命名为2 F 9。该株单抗细胞培养上清ELISA效价为1∶1 600,腹水效价为6.4×104,交叉反应结果表明2 F 9除了与测试的多株哈维氏弧菌发生免疫识别外,还与溶藻弧菌、麦氏弧菌和副溶血弧菌代表株发生免疫交叉反应,但不与拟态弧菌、鳗弧菌、嗜水气单胞菌代表株等发生交叉反应。用温和高碘酸氧化法对LPS处理后,GYC1108-1的LPS与2 F 9的反应部分减弱,免疫印迹结果表明2 F9识别LPS的类脂A抗原。用抑制ELISA法半定量检测GYC1108-1培养上清的LPS,测得培养48 h LPS释放量不超过39μg/mL,大大低于使鱼类致死所需的LPS的剂量。展开更多
Outbreaks of highly pathogenic avian influenza H5N1 virus have occurred in Vietnam since 2003. However, how people got avian H5N1 infection in Northern Vietnam is still unclear. We therefore performed a combination of...Outbreaks of highly pathogenic avian influenza H5N1 virus have occurred in Vietnam since 2003. However, how people got avian H5N1 infection in Northern Vietnam is still unclear. We therefore performed a combination of the serologic assays H5N1 ELISA and H5 western blot to detect anti-H5N1 specific antibodies. Sera samples of 149 subjects with suspected H5N1 infection from three provinces of Northern Vietnam were collected from September 2006 to March 2007. Our results indicated that this combinational assay showed high sensitivity (100%) and specificity (95%) when compared with hemagglutinin inhibition (HI) assay. Fifty-one sera samples (34.2%) contained specific antibodies against H5N1 viruses. Poultry raisers (32/77;41.6%) showed higher H5N1 infection rates than slaughterers (12/41;29.3%) and health care workers (7/31;22.6%). Contact history with sick or dead poultry in household or slaughter-house (p < 0.05) and lack of protective equipment use when in contact with dead poultry (p < 0.05) were risk factors found to be associated with H5N1 infection. In this study, we established an alternative serologic assay for H5N1 diagnosis, and we hereby present seroepidemiologic data of H5N1 infection in Northern Vietnam.展开更多
文摘以哈维氏弧菌GYC1108-1细菌颗粒性抗原免疫8周龄雌性BALB/c小鼠,5次免疫后取其脾细胞与SP2/0-Ag-14骨髓瘤细胞融合。用间接酶联免疫吸附试验(ELISA)对杂交瘤进行筛选,阳性克隆经3次亚克隆后,获得一株针对GYC1108-1脂多糖(LPS)的特异性单克隆抗体,命名为2 F 9。该株单抗细胞培养上清ELISA效价为1∶1 600,腹水效价为6.4×104,交叉反应结果表明2 F 9除了与测试的多株哈维氏弧菌发生免疫识别外,还与溶藻弧菌、麦氏弧菌和副溶血弧菌代表株发生免疫交叉反应,但不与拟态弧菌、鳗弧菌、嗜水气单胞菌代表株等发生交叉反应。用温和高碘酸氧化法对LPS处理后,GYC1108-1的LPS与2 F 9的反应部分减弱,免疫印迹结果表明2 F9识别LPS的类脂A抗原。用抑制ELISA法半定量检测GYC1108-1培养上清的LPS,测得培养48 h LPS释放量不超过39μg/mL,大大低于使鱼类致死所需的LPS的剂量。
文摘Outbreaks of highly pathogenic avian influenza H5N1 virus have occurred in Vietnam since 2003. However, how people got avian H5N1 infection in Northern Vietnam is still unclear. We therefore performed a combination of the serologic assays H5N1 ELISA and H5 western blot to detect anti-H5N1 specific antibodies. Sera samples of 149 subjects with suspected H5N1 infection from three provinces of Northern Vietnam were collected from September 2006 to March 2007. Our results indicated that this combinational assay showed high sensitivity (100%) and specificity (95%) when compared with hemagglutinin inhibition (HI) assay. Fifty-one sera samples (34.2%) contained specific antibodies against H5N1 viruses. Poultry raisers (32/77;41.6%) showed higher H5N1 infection rates than slaughterers (12/41;29.3%) and health care workers (7/31;22.6%). Contact history with sick or dead poultry in household or slaughter-house (p < 0.05) and lack of protective equipment use when in contact with dead poultry (p < 0.05) were risk factors found to be associated with H5N1 infection. In this study, we established an alternative serologic assay for H5N1 diagnosis, and we hereby present seroepidemiologic data of H5N1 infection in Northern Vietnam.
