NEW YORK – South Korean researchers reported on Wednesday that they have developed culture-free method for blood-based pathogen identification and antimicrobial susceptibility profiling that they said could hasten the treatment of patients with suspected sepsis by reducing testing times by 40 to 60 hours compared with typical antimicrobial susceptibility testing workflows used in hospitals.

纽约——韩国研究人员周三报告称，他们已经开发出无培养方法用于血液病原体鉴定和抗菌药物敏感性分析，他们说，与医院使用的典型抗菌药物敏感性测试工作流程相比，通过将测试时间缩短40至60小时，可以加速疑似败血症患者的治疗。

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In an article published in Nature, the researchers from Seoul National University and Seoul-based in vitro diagnostics firm QuantaMatrix said that they used a synthetic peptide to recover bacterial pathogens from whole-blood samples for identification and susceptibility testing, with results achieved in about 13 hours.

首尔国立大学和首尔体外诊断公司QuantaMatrix的研究人员在《自然》杂志上发表的一篇文章中表示，他们使用合成肽从全血样本中回收细菌病原体进行鉴定和药敏试验，结果在约13小时内取得。

Lead author Tae Hyun Kim said by email that QuantaMatrix, which was founded in 2010 as a Seoul National University spinout, is licensing the ultrarapid antimicrobial susceptibility testing (URAST) method with plans to develop a fully automated instrument with integrated pathogen detection, identification, and antimicrobial susceptibility profiling.

首席作者Tae Hyun Kim在电子邮件中表示，QuantaMatrix成立于2010年，是首尔国立大学的一家分支机构，它正在授权超快速抗菌药物敏感性测试（URAST）方法，计划开发一种全自动仪器，该仪器具有集成的病原体检测，鉴定和抗菌药物敏感性分析。

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The researchers said that they expect that the URAST system will 'completely transform the current standards of bacteremia diagnostics,' adding their methods have the potential to deliver phenotypic drug susceptibility information on the same day as the patient blood draw.

研究人员表示，他们预计URAST系统将“彻底改变当前的菌血症诊断标准”，并补充说，他们的方法有可能在患者抽血的同一天提供表型药物敏感性信息。

'With the potential to guide clinicians towards timely administration of optimal therapy, thereby saving many lives, our platform can also help deescalate the spread of antimicrobial resistance and sustain the lifespan of existing antibiotics,' they wrote.

他们写道：“我们的平台有可能指导临床医生及时给予最佳治疗，从而挽救许多生命，我们的平台还可以帮助减少抗菌药物耐药性的传播，并维持现有抗生素的寿命。”。

Kim said that the researchers aimed to develop testing methods that forgo blood culture because it is the most time-consuming process in antimicrobial susceptibility testing. He said that many hospitals also do not provide 24-hour lab services and their standard sepsis testing processes require manual steps that each introduce additional delays, further delaying test turnaround times beyond the critical treatment window for sepsis..

金说，研究人员旨在开发放弃血培养的检测方法，因为这是抗菌药物敏感性检测中最耗时的过程。他说，许多医院也不提供24小时实验室服务，他们的标准脓毒症检测过程需要手动步骤，每个步骤都会带来额外的延迟，进一步延迟脓毒症关键治疗窗口之外的检测周转时间。。

'Considering that patients' blood contains many growth inhibitors such as white blood cells and pre-administered antibiotics, we focused on approaches to directly isolate pathogens from whole blood,' he said.

他说：“考虑到患者血液中含有许多生长抑制剂，如白细胞和预先服用的抗生素，我们专注于从全血中直接分离病原体的方法。”。

Developing faster methods for detecting blood-borne infections and identifying their causes has been an enduring problem that other firms have also sought to address by eliminating the need to culture blood.

开发更快的方法来检测血源性感染并确定其原因一直是一个持久的问题，其他公司也试图通过消除血液培养的需要来解决这个问题。

T2 Biosystems has been a leader in the development of direct-from-blood sepsis tests with its T2Bacteria panel that is used for the detection of six common sepsis-causing pathogens within three to five hours. The firm has been developing a drug resistance panel with financial backing from the US Health and Human Services..

T2生物系统公司（T2 Biosystems）是直接从血脓毒症测试开发的领导者，其T2细菌小组用于在三到五个小时内检测六种常见的败血症病原体。该公司一直在开发一个由美国卫生与公众服务部资助的耐药性小组。。

German startup Noscendo recently began commercialization of a sequencing-based test, Disqver, that uses cell-free DNA in patient plasma samples, with an estimated 24 hours to receive definitive results.

德国初创公司Noscendo最近开始将基于测序的测试Disqver商业化，该测试在患者血浆样本中使用无细胞DNA，估计需要24小时才能获得明确的结果。

Meanwhile, Oxford Nanopore Technologies and Day Zero Diagnostics formed late last year a collaboration to develop a whole-genome sequencing-based clinical diagnostic system for the same-day identification and antimicrobial susceptibility profiling of bloodstream infections without the need for blood culture, while Siemens Healthineers has been developing a next-generation sequencing-based test to identify bacteria and fungi in a patient's bloodstream within six hours..

与此同时，牛津纳米孔技术公司（Oxford Nanopore Technologies）和Day Zero Diagnostics公司（Day Zero Diagnostics）于去年年底合作开发了一种基于全基因组测序的临床诊断系统，用于在不需要血培养的情况下对血流感染进行当天鉴定和抗菌药物敏感性分析，而西门子Healthineers公司（Siemens Healthineers）正在开发一种基于测序的下一代测试，以在六小时内识别患者血液中的细菌和真菌。。

Other firms including  Immunexpress, Inflammatix, Bluejay Diagnostics, Diasorin, and Cytovale have taken a different approach to improving sepsis response through rapid tests that are meant to quickly indicate which patients likely have sepsis or other severe infections or to guide patient triage.

包括Immunexpress，Inflammatix，Bluejay Diagnostics，Diasorin和Cytovale在内的其他公司已经采取了不同的方法，通过快速检测来改善败血症反应，这些检测旨在快速指示哪些患者可能患有败血症或其他严重感染，或者指导患者分类。

In the Nature article, the authors isolated bacterial pathogens from blood samples using magnetic nanoparticles with a synthetic version of human beta-2-glycoprotein 1 peptides which bind to a diverse array of bacteria species. Kim said that the team also evaluated the use of other immune response-related substances, including mannose-binding lectin proteins, but found that nanoparticles coated with the synthetic beta-2-glycoprotein 1 peptides had the highest binding affinity..

在《自然》杂志的文章中，作者使用磁性纳米粒子从血液样本中分离出细菌病原体，该纳米粒子具有与多种细菌物种结合的人β-2-糖蛋白1肽的合成版本。Kim说，该团队还评估了其他免疫反应相关物质的使用，包括甘露糖结合凝集素蛋白，但发现涂有合成β-2-糖蛋白1肽的纳米颗粒具有最高的结合亲和力。。

The enriched bacteria were identified using a multiplex assay with single-stranded DNA probes that were immobilized on the surface of shape-encoded microdisks with unique hole patterns for each target gene. Bacterial cells were lysed for genomic DNA extraction followed by amplification using nested PCR and biotin labeling.

使用单链DNA探针的多重测定法鉴定富集的细菌，所述单链DNA探针固定在shape编码的微盘的表面上，每个靶基因具有独特的孔模式。。

The researchers used the hole patterns and fluorescent dyes to aid gene-based identification of pathogens and achieved results within three hours..

研究人员使用孔模式和荧光染料来帮助基于基因的病原体鉴定，并在三小时内取得了结果。。

The authors reported that they also accelerated the antimicrobial susceptibility testing process by culturing the enriched bacteria in a purified medium. Under those conditions, rapidly dividing species such as Escherichia coli can reach the minimum cell count for URAST within four hours which allowed antimicrobial susceptibility testing with results in less than eight hours..

作者报告说，他们还通过在纯化培养基中培养富集的细菌来加速抗菌药物敏感性测试过程。在这些条件下，快速分裂的物种（如大肠杆菌）可以在四小时内达到尿毒症的最低细胞计数，从而可以在不到八小时内进行抗菌药物敏感性测试。。

To further reduce turnaround times, the researchers said that they evaluated URAST using custom-built 96-well chips that accommodate a low number of cellular inputs for minimum inhibitory concentration determination and susceptibility analysis.

为了进一步减少周转时间，研究人员表示，他们使用定制的96孔芯片评估了URAST，该芯片可容纳少量细胞输入，用于最小抑制浓度测定和敏感性分析。

The research team validated the URAST system in a 190-patient clinical study that paired the peptide-based enrichment methods with simultaneous tests for pathogen identification and susceptibility profiling. They reported that URAST identified bacterial pathogens with 100 percent concurrence with hospital-established testing protocols that used blood culture and matrix-assisted laser desorption-ionization time of flight (MALDI-TOF) mass spectrometry..

该研究团队在一项190名患者的临床研究中验证了URAST系统，该研究将基于肽的富集方法与病原体鉴定和药敏分析的同时测试配对。。。

The team also performed retrospective antimicrobial susceptibility testing on six clinical isolates and found that their testing methods performed with 94.9 percent agreement and a 5.1 percent minor error rate in comparison with broth microdilution disk diffusion. It said that their tests in the study delivered antimicrobial susceptibility results an average of 48 hours more quickly than conventional hospital workflows..

该团队还对六种临床分离株进行了回顾性抗菌药物敏感性测试，发现与肉汤微量稀释纸片扩散相比，他们的测试方法具有94.9%的一致性和5.1%的轻微错误率。它说，他们在研究中的测试比传统的医院工作流程平均快48小时。。

While the researchers tested for 36 bacteria species during their study, Kim noted that QuantaMatrix intends to expand the target range to at least 37 bacterial species and six fungi, so that the test that could be used for the detection and characterization of more than 95 percent of pathogens that cause sepsis.

虽然研究人员在研究期间测试了36种细菌，但金指出，QuantaMatrix打算将目标范围扩大到至少37种细菌和6种真菌，以便该测试可用于检测和表征95%以上引起败血症的病原体。

He said that the highly multiplexed identification assay also allows for the expansion of the gene list without significant increases in testing costs or processes, and the company hopes to add testing for 27 genes that are connected with antimicrobial susceptibility and resistance..

他说，高度多重的鉴定分析还可以在不显着增加测试成本或过程的情况下扩展基因列表，该公司希望增加对27个与抗菌药物敏感性和耐药性相关的基因的测试。。

Kim said that the team aims to build a system that would allow the seamless identification and characterization of pathogens within a blood sample with minimal involvement from clinicians so that the testing system can be implemented in clinical settings.

Kim说，该团队的目标是建立一个系统，可以在临床医生的最小参与下无缝识别和表征血液样本中的病原体，以便测试系统可以在临床环境中实施。

The Nature article notes that one of the patients in the clinical trial died during the testing period and a shorter turnaround time for testing may have helped healthcare providers to deliver optimal treatments and prevent renal failure. The patient had developed a fever 12 days after beginning chemotherapy for natural killer/T cell lymphoma and treatment began with the carbapenem antibiotic meropenem followed by Gilead's antifungal drug AmBisome (amphotericin B)..

《自然》杂志的文章指出，临床试验中的一名患者在测试期间死亡，测试的周转时间缩短可能有助于医疗保健提供者提供最佳治疗并预防肾衰竭。该患者在开始自然杀伤/T细胞淋巴瘤化疗12天后发烧，治疗开始于碳青霉烯类抗生素美罗培南，然后是吉利德的抗真菌药物AmBisome（两性霉素B）。。

Two days after treatment began, test results identified the cause of the infection as a carbapenem-resistant strain of Klebsiella pneumoniae.

治疗开始两天后，检测结果确定感染原因为耐碳青霉烯类肺炎克雷伯菌。

'We have been dedicated to AST research for over 10 years, dreaming of the day when our technology will reach the clinic and save many patients' lives,' Kim said. 'During the clinical pilot study, we witnessed numerous cases of sepsis patients passing away, which deepened our sense of responsibility towards this research.'.

金说：“我们致力于AST研究已有10多年，梦想着有一天我们的技术能够进入临床，挽救许多患者的生命。”在临床试验研究期间，我们目睹了许多脓毒症患者的死亡，这加深了我们对这项研究的责任感。”。