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评价产 IMP-27 摩根摩根菌的 NG-Test CARBA 5 侧向流检测试剂盒及其他…

Evaluation of the NG-Test CARBA 5 Lateral Flow Assay with an IMP-27-Producing Morganella morganii and Other .

机构信息

Department of Pathology and Immunology, Division of Laboratory and Genomic Medicine, Washington University School of Medicine, St. Louis, Missouri, USA.

Edison Family Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St. Louis, Missouri, USA.

出版信息

Microbiol Spectr. 2023 Jun 15;11(3):e0079323. doi: 10.1128/spectrum.00793-23. Epub 2023 May 18.

Abstract

An isolate of Morganella morganii (MMOR1) that tested susceptible to 3/4-generation cephalosporins and intermediate to meropenem was characterized as positive for NDM and IMP carbapenemases by NG-Test CARBA 5. Our objective was to further investigate this result, given the inconsistent susceptibility profile and unusual epidemiological profile for our region. The MMOR1 isolate was retested for antimicrobial susceptibilities and characterized for carbapenemase production. MMOR1 tested susceptible to ceftazidime, ceftriaxone, cefepime, aztreonam, and ertapenem, and intermediate to meropenem and imipenem. The isolate tested positive by carbapenem inactivation method (CIM) and CIM+EDTA (eCIM) testing, indicating metallo-β-lactamase production. The isolate tested negative for all carbapenemase genes on Xpert Carba-R, but positive for IMP on repeat testing of NG-Test CARBA 5. Whole-genome sequencing revealed MMOR1 contained , but no other carbapenemase genes. Additional testing with NG-Test CARBA 5 revealed a false-positive NDM band when the assay was overloaded with test inoculum. Supplementary isolates were tested with an overloaded inoculum ( = 6 M. morganii;  = 1 P. mirabilis;  = 1 IMP-27-producing ;  = 1 IMP-1-producing E. coli;  = 1 K. pneumoniae), and two non-carbapenemase-producing carbapenem non-susceptible M. morganii also generated a false-positive NDM band; though, this was not universal among this species. A dual IMP+/NDM+ M. morganii is an unusual result that should prompt additional investigation, especially in nonendemic regions and when the susceptibility profile is incompatible. IMP-27 is not detected by Xpert Carba-R but is variably detected by NG-Test CARBA 5. The microorganism inoculum used for NG-Test CARBA 5 must be carefully controlled for accurate results. The detection of carbapenemase-producing carbapenem-resistant Enterobacterales (CP-CRE) is an important function of the clinical microbiology laboratory, where positive identifications have immediate implications for infection control and surveillance strategies in the inpatient setting and can inform appropriate selection of therapy among the various novel anti-CP-CRE agents. NG-Test CARBA 5 is a relatively new lateral flow assay used for detection of carbapenemases in CP-CRE. Here, we describe the characterization of a Morganella morganii isolate that generated a false-positive NDM carbapenemase detection by this assay, and perform bacterial test inoculum experiments with additional isolates to further investigate a cause of false-positive results using the NG-Test CARBA 5. While a lateral flow assay like the NG-Test CARBA 5 is a very desirable test format for clinical laboratories, there are pitfalls to avoid when performing this test and interpreting results, including recognizing an overloaded test assay, which could lead to false-positive results.

摘要

一株对 3/4 代头孢菌素敏感、美罗培南中介的摩根摩根菌(MMOR1),通过 NG-Test CARBA 5 确认为 NDM 和 IMP 碳青霉烯酶阳性。鉴于我们地区不一致的药敏谱和不寻常的流行病学特征,我们的目标是进一步研究这一结果。对 MMOR1 分离株进行了抗菌药物敏感性检测,并对碳青霉烯酶的产生进行了鉴定。MMOR1 对头孢他啶、头孢曲松、头孢吡肟、氨曲南和厄他培南敏感,对美罗培南和亚胺培南中介。通过碳青霉烯失活法(CIM)和 CIM+EDTA(eCIM)检测,该分离株呈碳青霉烯酶阳性,表明产生金属β-内酰胺酶。Xpert Carba-R 检测所有碳青霉烯酶基因均为阴性,但 NG-Test CARBA 5 重复检测 IMP 阳性。全基因组测序显示,MMOR1 含有 ,但没有其他碳青霉烯酶基因。用 NG-Test CARBA 5 进行额外检测时,当检测接种物过载时,会出现假阳性 NDM 带。用过量接种物( = 6 株摩根摩根菌;  = 1 株奇异变形杆菌;  = 1 株产 IMP-27 ;  = 1 株产 IMP-1 ;  = 1 株产 K. pneumoniae)进行了补充检测,两种非碳青霉烯酶产生的碳青霉烯类不敏感的摩根摩根菌也产生了假阳性 NDM 带;尽管在该物种中并非普遍存在。一个双重 IMP+/NDM+摩根摩根菌是一个不寻常的结果,应提示进行进一步调查,特别是在非流行地区和药敏谱不兼容时。Xpert Carba-R 无法检测到 IMP-27,但 NG-Test CARBA 5 可检测到。用于 NG-Test CARBA 5 的微生物接种物必须仔细控制,以获得准确的结果。检测产碳青霉烯酶的碳青霉烯类耐药肠杆菌科(CP-CRE)是临床微生物学实验室的一项重要功能,阳性鉴定立即对住院患者的感染控制和监测策略产生影响,并能为各种新型抗 CP-CRE 药物的选择提供信息。NG-Test CARBA 5 是一种用于检测 CP-CRE 中碳青霉烯酶的新型侧向流动检测法。在这里,我们描述了一种摩根摩根菌分离株的特征,该分离株通过该检测方法产生了假阳性 NDM 碳青霉烯酶检测,并用其他分离株进行细菌测试接种物实验,进一步研究了使用 NG-Test CARBA 5 产生假阳性结果的原因。虽然像 NG-Test CARBA 5 这样的侧向流动检测法是临床实验室非常理想的检测方法,但在进行该检测和解释结果时存在一些需要避免的陷阱,包括识别过载的检测试验,这可能导致假阳性结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f5c9/10269506/aa8719315d70/spectrum.00793-23-f001.jpg

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