首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Unstable Expression and Thermal Instability of a Species-Specific Cell Surface Epitope Associated with a 66-Kilodalton Antigen Recognized by Monoclonal Antibody EM-7G1 within Serotypes of Listeria monocytogenes Grown in Nonselective and Selective Broths
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Unstable Expression and Thermal Instability of a Species-Specific Cell Surface Epitope Associated with a 66-Kilodalton Antigen Recognized by Monoclonal Antibody EM-7G1 within Serotypes of Listeria monocytogenes Grown in Nonselective and Selective Broths

机译:在非选择性和选择性培养液中生长的单核细胞增生李斯特菌血清型中与单克隆抗体EM-7G1识别的66-Kaldalton抗原相关的物种特异性细胞表面表位的不稳定表达和热不稳定性

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摘要

Conditions that resulted in unstable expression and heat instability of a cell surface epitope associated with a 66-kDa antigen in Listeria monocytogenes serotypes were identified with the probe monoclonal antibody (MAb) EM-7G1 in an enzyme-linked immunosorbent assay. This epitope appeared to be absent in three serotypes (serotypes 3b, 4a, and 4c), which did not react with MAb EM-7G1 irrespective of the enrichment broth tested. The remaining 10 serotypes were detected by MAb EM-7G1 only when cells were grown in nonselective brain heart infusion broth (BHI) or selective Listeria enrichment broth (LEB). When cells were grown in Listeria repair broth (LRB), only 6 of the 13 serotypes were detected by MAb EM-7G1, and recognition of serogroup 4 was completely lost. None of the 13 serotypes was detected by MAb EM-7G1 when cells were grown in two other commonly used Listeria-selective media, UVM1 broth and Fraser broth (FRB), indicating that possible loss of epitope expression occurred under these conditions. MAb EM-7G1 maintained species specificity without cross-reacting with live or heat-killed cells of six other Listeria spp. (Listeria ivanovii, Listeria innocua, Listeria seeligeri, Listeria welshimeri, Listeria grayi, and Listeria murrayi) irrespective of the enrichment conditions tested. Due to heat instability of the cell surface epitope when it was exposed to 80 or 100°C for 20 min, MAb EM-7G1 is suitable for detection of live cells of L. monocytogenes in BHI or LEB but not in LRB, UVM1, or FRB enrichment medium.
机译:在酶联免疫吸附试验中,用探针单克隆抗体(MAb)EM-7G1鉴定了导致单核细胞增生李斯特菌血清型中与66 kDa抗原相关的细胞表面表位表达不稳定和热不稳定性的条件。该表位似乎不存在于三种血清型(血清型3b,4a和4c)中,无论测试的富集肉汤如何,它们都不与MAb EM-7G1反应。仅当细胞在非选择性脑心脏输液肉汤(BHI)或选择性李斯特菌富集肉汤(LEB)中生长时,才通过MAb EM-7G1检测剩余的10种血清型。当细胞在李斯特菌修复肉汤(LRB)中生长时,MAb EM-7G1仅检测到13种血清型中的6种,并且完全失去了对4型血清型的识别。当细胞在另外两种常用的利斯特氏菌选择性培养基UVM1肉汤和Fraser肉汤(FRB)中生长时,MAb EM-7G1未检测到13种血清型,表明在这些条件下发生了表位表达的可能丧失。单抗EM-7G1保持物种特异性,而不会与其他六个李斯特菌属物种的活细胞或热灭活细胞发生交叉反应。 (伊万氏李斯特菌,无病李斯特菌,Seeligeri李斯特菌,welshimeri李斯特菌,灰色李斯特菌和murrayi李斯特菌),无论测试的富集条件如何。由于将其暴露于80或100°C 20分钟时细胞表面表位的热不稳定性,MAb EM-7G1适合检测BHI或LEB中单核细胞增生李斯特菌的活细胞,但不适用于LRB,UVM1或FRB富集培养基。

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