荧光素酶报告分枝杆菌噬菌体(TM4::GeNL)能够快速评估脓肿分枝杆菌复合体的药物敏感性和诱导大环内酯类药物耐药性。

IF 5.4 2区 医学 Q1 MICROBIOLOGY
Journal of Clinical Microbiology Pub Date : 2025-09-10 Epub Date: 2025-08-20 DOI:10.1128/jcm.00841-25
Saranathan Rajagopalan, Lahari Das, Donna J Kohlerschmidt, Amy K Rourke, Salika M Shakir, Michelle H Larsen, Max R O'Donnell, Wendy A Szymczak, Vincent E Escuyer, Phyu M Thwe, William R Jacobs
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引用次数: 0

摘要

由于对抗结核药物、碳青霉烯类药物、氟喹诺酮类药物和四环素具有高度耐药性,脓肿分枝杆菌(MAB)感染的治疗具有挑战性。克拉霉素和阿米卡星因其优越的疗效被认为是MAB治疗的基础药物;然而,由于诱导大环内酯类耐药,评估克拉霉素敏感性通常需要7至14天。采用TM4::GeNL建立48 h荧光素酶报告型分枝噬菌体药敏试验(LRM-DST)方法评价单克隆抗体的药敏。在这项原理验证研究中,我们使用TM4::GeNL和Sensititre RAPMYCO2板对26株单抗临床分离株进行了DST检测。进行基因组测序以鉴定MAB亚种,并了解内在和获得性耐药机制。我们通过Sensititre和LRM-DST检测到12/26(46%)的克拉霉素耐药。所有耐克拉霉素分离株均携带全长诱导型红霉素核糖体甲基化酶-41 [erm(41)]基因,无rrl (23S rRNA)突变。6/26株(23%)菌株对阿米卡星耐药或中间耐药。大肠杆菌148a > G位点的rrs (16S rRNA)基因突变导致4株菌株对阿米卡星产生耐药性;然而,在两株阿米卡星中等耐药菌株中未发现突变。LRM-DST结果与克拉霉素(kappa系数1.0,95%可信区间[CI] 0.84 ~ 1.0)和阿米卡星(kappa系数0.752,95%可信区间[CI] 0.43 ~ 1.0)的Sensititre DST结果的一致性分别为100%和92.3%。此外,贝达喹啉、莫西沙星、亚胺培南、利奈唑胺和头孢西丁的LRM-DST结果与Sensititre DST具有良好的相关性。我们的研究结果表明,LRM-DST可以在48小时内提供可靠的表型MAB-DST信息,以便及时进行临床管理。脓分枝杆菌(MAB)是一种臭名昭著的人类病原体,可引起囊性纤维化和免疫功能低下患者的严重感染。单克隆抗体的治疗选择非常有限,因为它们通过内在和获得性耐药机制对多种药物产生耐药性。虽然大环内酯类和阿米卡星是对抗单抗感染的关键药物,但新出现的耐药性正在损害它们的疗效。目前基于生长的药敏试验(DST)方法由于存在诱导型大环内酯类耐药,需要7 ~ 14天才能确定克拉霉素的药敏,其他药物需要3 ~ 5天。我们开发了一种新的基于荧光素酶报告型分枝噬菌体(LRM)的表型DST方法,使用TM4::GeNL基于代谢抑制而不是生长来评估MAB药物敏感性。LRM-DST显着将DST周转时间缩短至48小时,并提供MAB对药物的敏感性快速评估,包括诱导大环内酯类耐药,从而加快治疗过程并改善患者预后。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Luciferase reporter mycobacteriophage (TM4::GeNL) enables rapid assessment of drug susceptibilities and inducible macrolide resistance in Mycobacterium abscessus complex.

Mycobacterium abscessus (MAB) infections are challenging to treat due to high-level resistance to anti-tuberculosis drugs, carbapenems, fluoroquinolones, and tetracyclines. Clarithromycin and amikacin are considered cornerstone drugs for MAB treatment due to their superior efficacy; however, assessing clarithromycin susceptibility typically takes 7 to 14 days because of inducible macrolide resistance. We developed a 48 h luciferase reporter mycobacteriophage drug susceptibility testing (LRM-DST) assay using TM4::GeNL to evaluate MAB drug susceptibility. For this proof-of-principle study, we performed DST on 26 MAB clinical isolates using TM4::GeNL and Sensititre RAPMYCO2 plates. Genome sequencing was performed to identify the MAB sub-species and for understanding intrinsic and acquired drug resistance mechanisms. We detected clarithromycin resistance in 12/26 (46%) by both Sensititre and LRM-DST. All clarithromycin-resistant isolates carried full-length inducible erythromycin ribosomal methylase-41 [erm(41)] gene, and none had rrl (23S rRNA) mutations. Amikacin resistance or intermediate amikacin resistance was detected in 6/26 (23%) isolates. Mutations in the rrs (16S rRNA) gene at positions 1375A > G (E. coli 1408A > G) conferred amikacin resistance in four isolates; however, no mutations were identified in two of the amikacin intermediate-resistant isolates. LRM-DST results were in 100% and 92.3% concordance with Sensititre DST results of clarithromycin (kappa coefficient, 1.0; 95% confidence interval [CI], 0.84 to 1.0) and amikacin (kappa coefficient, 0.752; 95% confidence interval [CI], 0.43 to 1.0). In addition, LRM-DST results for bedaquiline, moxifloxacin, imipenem, linezolid, and cefoxitin correlated well with Sensititre DST. Our findings suggest that LRM-DST can provide reliable phenotypic MAB-DST information in 48 h for prompt clinical management.IMPORTANCEMycobacterium abscessus (MAB) is a notorious human pathogen causing severe infections in individuals with cystic fibrosis and immunocompromised patients. Treatment options for MAB are very limited as they are resistant to multiple drugs through intrinsic and acquired resistance mechanisms. While macrolides and amikacin are the key drugs in the fight against MAB infections, emerging drug resistance is compromising their efficacy. Current growth-based drug susceptibility testing (DST) method takes 7 to 14 days to identify clarithromycin susceptibility due to the presence of inducible macrolide resistance and 3 to 5 days for other drugs. We developed a novel luciferase reporter mycobacteriophage (LRM) based phenotypic DST method using TM4::GeNL to assess MAB drug susceptibility based on metabolic inhibition rather than growth. LRM-DST significantly reduces the DST turnaround time to 48 h and provides rapid assessment of MAB susceptibility to drugs, including inducible macrolide resistance, thereby accelerating the treatment process and improving patient outcomes.

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来源期刊
Journal of Clinical Microbiology
Journal of Clinical Microbiology 医学-微生物学
CiteScore
17.10
自引率
4.30%
发文量
347
审稿时长
3 months
期刊介绍: The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.
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