Development of the MasSpec Pen Technology for Rapid and Accurate Identification of Pediatric Infections

摘要：骨关节感染（OAI）是不成比例地影响儿童的常见疾病， 这可能导致严重的并发症，包括运动功能丧失和骨畸形。快速准确诊断 的OAI病原体是至关重要的，以允许选择特定的和有针对性的治疗方案，并改善结果， 儿科患者。然而，在临床实践中，患者经常开始使用广谱抗生素， 通过培养或聚合酶链反应进行鉴定，这可能需要几天才能完成。此外，高 培养阴性OAI的发生率和抗生素耐药菌株的流行率增加， 对OAI的挑战同时，使用多种抗生素的非特异性治疗方案可导致多种短期和长期- 长期不良影响。在这里，我们建议开发和测试MasSpec Pen（MSPen）技术作为一种创新方法 用于直接从临床标本中快速准确地鉴定OAI。我们开发了MSPen作为一个易于使用的 手持式设备集成到高性能质谱仪中，用于快速（<20秒）检测丰富的代谢产物 直接从生物样本中提取的图谱。现在，我们假设MSPen可以发展成为一种强大的技术， 用于从OAI临床样本中快速、直接和准确地鉴定病原体，从而推进 通过允许选择特异性和靶向治疗来治疗儿科患者。通过Livia S. Eberlin（德克萨斯大学奥斯汀分校化学系），质谱专家， MSPen技术，Sarmistha B博士。Hauger（戴尔医学院儿科传染病主任）和Lindsey博士 柯克帕特里克（传染病系，印第安纳州大学医学院），其医疗实践致力于 儿科感染的治疗，我们建议通过执行以下具体目标来测试我们的假设： 目标1.优化MSPen检测试剂盒，用于儿科OAI临床标本和相关培养物的分子分析 分离株MSPen在复杂生物样品的直接分析中提供了变革性的分子检测能力， 样品沿着具有对常规临床使用有吸引力的操作特征。我们将改进设备和方法， 优化其分析性能，用于OAI临床标本和相应培养物的直接分子分析 分离株目标2.确定MSPen用于儿科OAI病原体鉴定的诊断性能 样品MSPen有潜力提供分子信息，以推动快速准确的治疗决策- 制作。我们将分析前瞻性收集的儿科OAI样本，包括临床相关的 细菌菌株将开发统计分类器，以使用临床试验将细菌病原体分类到分类学水平。 微生物学结果作为金标准。目标3.评价MSPen鉴定抗生素的能力 耐药菌株。OAI儿科患者中抗生素耐药菌的发生率增加 揭示了对准确鉴定抗生素敏感性的迫切需要。我们将调查MSPen的能力 鉴定临床样本中的抗生素耐药菌和敏感菌。将开发统计分类器， 鉴定抗性或敏感S.金黄色葡萄球菌，使用临床微生物学结果作为金标准。

ABSTRACT: Osteoarticular infections (OAI) are prevalent medical conditions that disproportionately affect children, which can lead to serious complications including loss of motor function and bone deformity. Rapid and accurate diagnosis of OAI etiologic agents are critical to allow selection of specific and targeted treatment options and improve outcomes for pediatric patients. Yet, in clinical practice, patients are often started on broad spectrum antibiotics pending pathogen identification by culture or polymerase chain reaction, which can take several days for completion. In addition, the high rates of culture-negative OAI and the increased prevalence of antibiotic resistant strains make therapeutic decisions uniquely challenging for OAI. Meanwhile, unspecific therapy regimens with multiple antibiotics can lead to multiple short- and long- term adverse effects. Here, we propose to develop and test the MasSpec Pen (MSPen) technology as an innovative method for rapid and accurate identification of OAI directly from clinical specimens. We developed the MSPen as an easy-to-use handheld device integrated to a high-performance mass spectrometer for rapid (<20 seconds) detection of rich metabolic profiles directly from biological samples. Now, we hypothesize that the MSPen can be developed as a powerful technology for rapid, direct, and accurate identification of etiologic agents from OAI clinical samples, thus advancing treatment for pediatric patients by allowing selection of specific and targeted therapy. Through a collaboration between Prof. Livia S. Eberlin (Department of Chemistry, The University of Texas at Austin), expert in mass spectrometry and developer of the MSPen technology, Dr. Sarmistha B. Hauger (Chief of Pediatric Infectious Disease, Dell Medical School) and Dr. Lindsey Kirkpatrick (Infectious Disease Faculty, Indiana University School of Medicine), whose medical practices are dedicated to the treatment of pediatric infections, we propose to test our hypothesis by carrying out the following specific aims: Aim 1. Optimize the MSPen assay for molecular analysis of pediatric OAI clinical specimens and associated culture isolates. The MSPen provides transformative molecular detection capabilities in the direct analysis of complex biological samples along with operational features that are attractive for routine clinical use. We will refine the device and method to optimize its analytical performance for direct molecular analysis of OAI clinical specimens and corresponding culture isolates. Aim 2. Determine the diagnostic performance of the MSPen for pathogen identification in pediatric OAI samples. The MSPen has the potential to provide molecular information to drive rapid and accurate therapeutic decision- making. We will analyze pediatric OAI samples prospectively collected for our study encompassing clinically-relevant bacteria strains. Statistical classifiers will be developed to classify bacterial pathogens into taxonomical levels using clinical microbiology results as gold-standard. Aim 3. Evaluate the capabilities of the MSPen for the identification of antibiotic resistant bacterial strains. The increasing occurrence of antibiotic resistant bacteria in pediatric patients presenting OAI reveals a critical need for accurate identification of antibiotic susceptibility. We will investigate the capability of the MSPen in identifying antibiotic resistant and susceptible bacteria in clinical samples. Statistical classifiers will be developed to identify resistant or susceptible S. aureus, using clinical microbiology results as gold-standard.