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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Enzyme-linked immunosorbent assay using a recombinant baculovirus-expressed Bacillus anthracis protective antigen (PA): measurement of human anti-PA antibodies.
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Enzyme-linked immunosorbent assay using a recombinant baculovirus-expressed Bacillus anthracis protective antigen (PA): measurement of human anti-PA antibodies.

机译：使用重组杆状病毒表达的炭疽芽孢杆菌保护性抗原（PA）的酶联免疫吸附测定：人抗PA抗体的测量。

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We developed an antigen capture enzyme-linked immunosorbent assay (ELISA) which does not require purified protective antigen (PA) for detection of human antibodies to Bacillus anthracis PA. Lysates of Spodoptera frugiperda (Sf-9) cells infected with recombinant baculovirus containing the PA gene were used as the source of PA to develop the ELISA. Recombinant PA from crude Sf-9 cell lysates or PA purified from B. anthracis Sterne strain was captured by an anti-PA monoclonal antibody coated onto microtiter plates. We demonstrated that human serum antibody titers to PA were identical in the ELISA whether we used crude Sf-9 cell lysates containing recombinant baculovirus-expressed PA or purified Sterne PA. Finally, false-positive results observed in a direct ELISA were eliminated with this antigen capture ELISA. Thus, the antigen capture ELISA with crude preparations of baculovirus-expressed PA is reliable, safe, and inexpensive for determining anti-PA antibody levels in human sera.
机译：我们开发了一种无需进行纯化的保护性抗原（PA）即可检测抗炭疽芽孢杆菌PA的人类抗体的抗原捕获酶联免疫吸附测定（ELISA）。用含有PA基因的重组杆状病毒感染的斜纹夜蛾（Sf-9）细胞裂解液用作PA的来源，以开发ELISA。通过包被在微量滴定板上的抗PA单克隆抗体捕获来自粗Sf-9细胞裂解物的重组PA或从炭疽芽孢杆菌Sterne菌株纯化的PA。我们证明，无论我们使用含有重组杆状病毒表达的PA还是纯化的Sterne PA的Sf-9细胞粗裂解物，ELISA中人血清对PA的抗体滴度都是相同的。最后，用这种抗原捕获ELISA消除了直接ELISA中观察到的假阳性结果。因此，用杆状病毒表达的PA的粗制品进行抗原捕获ELISA对于确定人血清中的抗PA抗体水平是可靠，安全和便宜的。

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