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    • A Proof-of-principle Study for the Point-of-care Detection of ESBL (CTX-M) by NG-Test® CTX-M MULTI Lateral Flow Assay in Urine Samples Using a Simplified Method for Use in a Resource-Limited Setting

    Dennis Nurjadi,Arnaud Chalin,Susanne Hauswaldt,Linus Olson,Mattias Larsson, Ase Ostholm,Thirumalaisamy P. Velavan,Sebastien Boutin,Jan Rupp,Lennart E. Nilsson,Hakan Hanberger

    JAC-Antimicrobial Resistance（2024）

    Department of Infectious Diseases and Microbiology | NG Biotech | Karolinska Inst | Dept Infect Dis Reg Ostergotland | Vietnamese German Ctr Med Res VGCARE | Univ Lubeck | Linkoping Univ | Training & Res Acad Collaborat TRAC

    Cited 0|Views4
    Abstract
    Background: The rise of extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-E) in low- and middle-income countries limits treatment options, leading to the frequent use of broad-spectrum antibiotics. Reducing time-to-result for a urinary infection can facilitate correct antibiotic treatment and support antimicrobial and diagnostic stewardship measures. This study compared two simplified enrichment methods for detecting CTX-M directly from urine specimens. Methods: Two enrichment methods, namely centrifugation of 2 mL urine and filtration of 1 mL urine using the DirecTool adaptor, were compared using 20 culture-positive urine samples (20 suspected ESBL-E and 20 non-ESBL-E). CTX-M production was detected using a lateral flow assay (LFA), NG-Test (R) CTX-MMULTI. The presence of bla(CTX-M) genes was confirmed by whole-genome sequencing (WGS). Results: The results of both enrichment methods were identical, with a sensitivity of 87.5% and a specificity of 100%. In 19/20 (95%) of the urine samples, the results of the CTX-M LFA were identical with the phenotypic confirmation and WGS. Both methods could detect ESBL-E bacteriuria with >= 10(4) cfu/mL. All ESBL-E-negative samples were identified accurately. Both enrichment methods yielded negative results in one ESBL-E-positive (CTX-M-15) sample despite phenotypic and genotypic confirmation of ESBL production. High leukocyte count (>500 cells/mu L), the presence of boric acid or polymicrobial samples did not appear to impact the performance of both enrichment methods. Conclusions: Our study underscores the feasibility of directly detecting CTX-M in urine. Simplified enrichment methods, particularly with a filtration kit, enhance the assay's practicality, rendering it suitable for use in primary care, emergency departments or remote laboratories without sophisticated equipment.
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    【要点】：本研究验证了一种在资源有限环境下使用简化方法，通过尿液样本中的NG-Test® CTX-M MULTI侧流免疫层析检测法快速检测ESBL（CTX-M）的可行性，旨在缩短尿路感染诊断时间，从而优化抗生素治疗。
    

    【方法】：研究比较了两种简化富集方法——尿样离心和尿样通过DirecTool适配器过滤，使用侧流免疫层析检测法（LFA）检测CTX-M的产生，并以全基因组测序（WGS）确认bla(CTX-M)基因的存在。
    

    【实验】：实验使用了20份培养阳性的尿液样本（怀疑为ESBL-E的20份样本和20份非ESBL-E样本）。两种富集方法均显示出87.5%的灵敏度和100%的特异性，且95%的样本中LFA结果与表型确认和WGS结果一致，能够检测到菌落形成单位每毫升大于等于10^4的ESBL-E菌尿。所有ESBL-E阴性样本均被准确识别。在高白细胞计数、硼酸存在或多微生物样本的情况下，两种方法性能不受影响。
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