Moshgabadi Noushin, Galli Rick A, Daly Amelia C, Ko Sze Mun Shirley, Westgard Tayla E, Bulpitt Ashley F, Shackleton Christopher R
bioLytical Laboratories Inc., 1108-13351 Commerce Parkway, Richmond, BC V6V 2X7, Canada.
bioLytical Laboratories Inc., 1108-13351 Commerce Parkway, Richmond, BC V6V 2X7, Canada.
J Clin Virol. 2015 Oct;71:67-72. doi: 10.1016/j.jcv.2015.08.005. Epub 2015 Aug 13.
Anti-HIV-1 IgM antibody is an important immunoassay target for early HIV antibody detection.
The objective of this study is to determine if the early HIV antibody sensitivity of the 60s INSTI test is due to detection of anti-HIV-1 IgM in addition to IgG.
To demonstrate HIV gp41 IgM antibody capture by the INSTI HIV-1 gp41 recombinant antigen, an HIV-IgM ELISA was conducted with commercial HIV-1 seroconversion samples. To demonstrate that the INSTI dye-labelled Protein A-based colour developer (CD) has affinity to human IgM, commercial preparations of purified human immunoglobulins (IgM, IgD, IgA, IgE, and IgG) were blotted onto nitrocellulose (NC) and probed with the CD to observe spot development. To determine that INSTI is able to detect anti-HIV-1 IgM antibody, early seroconversion samples, were tested for reduced INSTI test spot intensity following IgM removal.
The gp41-based HIV-IgM ELISA results for 6 early seroconversion samples that were INSTI positive determined that the assay signal was due to anti-HIV-1 IgM antibody capture by the immobilised gp41 antigen. The dye-labelled Protein-A used in the INSTI CD produced distinct spots for purified IgM, IgA, and IgG blotted on the NC membrane. Following IgM removal from 21HIV-1 positive seroconversion samples with known or undetermined anti-HIV-1 IgM levels that were western blot negative or indeterminate, all samples had significantly reduced INSTI test spot intensity.
The INSTI HIV-1/HIV-2 Antibody Test is shown to detect anti-HIV-1 IgM antibodies in early HIV infection which enhances its utility in early HIV diagnosis.
抗HIV-1 IgM抗体是早期HIV抗体检测的重要免疫测定靶点。
本研究的目的是确定60s整合酶链转移抑制剂（INSTI）检测对早期HIV抗体的敏感性是否归因于除IgG外还检测到抗HIV-1 IgM。
为了证明INSTI HIV-1 gp41重组抗原捕获HIV gp41 IgM抗体，使用商业HIV-1血清转化样本进行了HIV-IgM酶联免疫吸附测定（ELISA）。为了证明INSTI基于染料标记蛋白A的显色剂（CD）对人IgM有亲和力，将纯化的人免疫球蛋白（IgM、IgD、IgA、IgE和IgG）商业制剂点样到硝酸纤维素膜（NC）上，并用显色剂进行检测以观察斑点显色情况。为了确定INSTI能够检测抗HIV-1 IgM抗体，对早期血清转化样本在去除IgM后检测INSTI检测斑点强度是否降低。
对6份INSTI检测呈阳性的早期血清转化样本进行基于gp41的HIV-IgM ELISA检测，结果表明检测信号归因于固定化gp41抗原捕获抗HIV-1 IgM抗体。INSTI显色剂中使用的染料标记蛋白A在点样于NC膜上的纯化IgM、IgA和IgG上产生了明显的斑点。在用已知或未知抗HIV-1 IgM水平且蛋白质印迹法检测为阴性或不确定的21份HIV-1阳性血清转化样本中去除IgM后，所有样本的INSTI检测斑点强度均显著降低。
INSTI HIV-1/HIV-2抗体检测显示能够在早期HIV感染中检测抗HIV-1 IgM抗体，这增强了其在早期HIV诊断中的效用。
