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在酶联免疫吸附测定中使用伯氏疏螺旋体和嗜吞噬细胞无形体的重组抗原来检测白尾鹿体内的抗体。

Use of recombinant antigens of Borrelia burgdorferi and Anaplasma phagocytophilum in enzyme-linked immunosorbent assays to detect antibodies in white-tailed deer.

作者信息

Magnarelli Louis A, IJdo Jacob W, Ramakrishnan Uma, Henderson David W, Stafford Kirby C, Fikrig Erol

机构信息

Department of Entomology and Forestry and Horticulture, The Connecticut Agricultural Experiment Station, P.O. Box 1106, New Haven, Connecticut 06504, USA.

出版信息

J Wildl Dis. 2004 Apr;40(2):249-58. doi: 10.7589/0090-3558-40.2.249.

Abstract

Serum samples obtained from white-tailed deer (Odocoileus virginianus) in Connecticut (n=218) and South Carolina (n=20) (USA) during the period 1992-2002 were analyzed for antibodies to whole-cell or recombinant antigens (i.e., fusion proteins) of Borrelia burgdorferi sensu stricto and Anaplasma phagocytophilum, etiologic agents of Lyme borreliosis and granulocytic ehrlichiosis, respectively. In enzyme-linked immunosorbent assays (ELISAs) with whole-cell B. burgdorferi, the overall seropositivity rate for Connecticut (53%) exceeded that for South Carolina (30%). In separate tests of seven recombinant antigens of B. burgdorferi by an ELISA, seroprevalence for the VlsE antigen was highest (48%) in Connecticut followed by outer surface protein (OspF) (21%), whereas serum reactivities to the protein (p) 41-G antigen (55%) and VlsE (25%) were most frequent for South Carolina sera. In analyses for antibodies to the recombinant protein (p) 44 antigen of A. phagocytophilum, seroprevalences of 52% and 25% were recorded for Connecticut and South Carolina samples, respectively. These findings paralleled those determined by indirect fluorescent antibody staining methods with whole cells (43% and 30%). Moreover, there was good agreement (74%) in results of Western blot analyses and an ELISA when a subset of 39 sera was screened with whole-cell or recombinant p44 antigens of A. phagocytophilum. An ELISA with highly specific recombinant VlsE or p44 antigens can be used in conjunction with other antibody tests to determine whether deer living in different regions of eastern United States were exposed to B. burgdorferi or A. phagocytophilum.

摘要

1992年至2002年期间,从美国康涅狄格州(n = 218)和南卡罗来纳州(n = 20)的白尾鹿(弗吉尼亚鹿)采集血清样本,分析其针对伯氏疏螺旋体狭义种和嗜吞噬细胞无形体全细胞或重组抗原(即融合蛋白)的抗体,这两种病原体分别是莱姆病和粒细胞埃立克体病的病原体。在针对伯氏疏螺旋体全细胞的酶联免疫吸附测定(ELISA)中,康涅狄格州的总体血清阳性率(53%)超过南卡罗来纳州(30%)。通过ELISA对伯氏疏螺旋体的七种重组抗原进行单独检测时,康涅狄格州VlsE抗原的血清阳性率最高(48%),其次是外表面蛋白(OspF)(21%),而南卡罗来纳州血清对41-G抗原(p41-G)(55%)和VlsE(25%)的血清反应最为常见。在分析针对嗜吞噬细胞无形体重组蛋白(p)44抗原的抗体时,康涅狄格州和南卡罗来纳州样本的血清阳性率分别为52%和25%。这些发现与采用全细胞间接荧光抗体染色法得出的结果(43%和30%)相似。此外,当用嗜吞噬细胞无形体的全细胞或重组p44抗原对39份血清进行筛选时,蛋白质印迹分析和ELISA结果的一致性良好(74%)。采用具有高度特异性的重组VlsE或p44抗原进行ELISA,可与其他抗体检测方法结合使用,以确定生活在美国东部不同地区的鹿是否接触过伯氏疏螺旋体或嗜吞噬细胞无形体。

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