摘要

移植粘菌素抗性基因mcr-1最近在从广泛来源分离的细菌中发现并迅速传播，这破坏了我们治疗细菌感染和威胁人类健康和安全的能力。为了防止粘菌素抗性的进一步转移，需要实用且可靠的含mcr-1细菌的方法。在该研究中，开发了针对MCR-1的标准和新型多克隆和单克隆抗体（mAb）。在9种mAb中，3种是MCR-1特异性的，6种是与MCR-1和MCR-2交叉反应。使用多克隆抗体作为捕获剂和mAb MCR-1-7作为检测器建立夹心酶联免疫吸附测定（ELISA）。对于MCR-1，该测定的检测限为0.01ng / mL，对于缓冲液中的MCR-2，检测限为0.1ng / mL，变异系数（CV）小于15％。当应用于碎牛肉，鸡肉和猪肉时，该ELISA鉴定了接种少于0.4cfu / g肉的样品，证明其对复杂食物基质的强耐受性。据我们所知，这是第一个针对MCR-1和MCR-2开发的免疫分析。它应该有助于迅速和可靠地筛选受质粒携带的粘菌素抗性细菌污染的肉类样品，从而降低人类食源性感染的风险，可能没有抗生素治疗方案。

The recent discovery and rapid spread of mobile colistin-resistant gene, mcr-1, among bacteria isolated from a broad range of sources is undermining our ability to treat bacterial infections and threatening human health and safety. To prevent further transfer of colistin resistance, practical and reliable methods for mcr-1-containing bacteria are need. In this study, standards and novel polyclonal and monoclonal antibodies (mAbs) against MCR-1 were developed. Among nine mAbs, three were MCR-1 specific and six cross-reacted with both MCR-1 and MCR-2. A sandwich enzyme-linked immunosorbent assay (ELISA) was established using the polyclonal antibody as a capturer and the mAb MCR-1-7 as a detector. The assay had a limit of detection of 0.01 ng/mL for MCR-1 and 0.1 ng/mL for MCR-2 in buffer with coefficients of variation (CV) less than 15%. When applied to ground beef, chicken and pork, this ELISA identified samples inoculated with less than 0.4 cfu/g of meat, demonstrating its strong tolerance to complex food matrices. To our knowledge, this is the first immunoassay developed for MCR-1 and MCR-2. It should be useful for prompt and reliable screening of meat samples contaminated with plasmid-borne colistin-resistant bacteria, thus reducing human risk of foodborne infections with possibly no antibiotic treatment options.

https://www.ncbi.nlm.nih.gov/pubmed/30425266