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首页> 外文期刊>The Laryngoscope: A Medical Journal for Clinical and Research Contributions in Otolaryngology, Head and Neck Medicine and Surgery, Facial Plastic and Reconstructive Surgery .. >Improving patient care via development of a protein-based diagnostic test for microbe-specific detection of chronic rhinosinusitis
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Improving patient care via development of a protein-based diagnostic test for microbe-specific detection of chronic rhinosinusitis

机译：通过开发基于蛋白质的诊断测试来改善慢性鼻鼻窦炎的微生物特异性检测，从而改善患者护理

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Objectives/Hypothesis The hypothesis is that signature bacterial proteins can be identified in sinus secretions via high-throughput, proteomic based techniques. Nontypeable Haemophilus influenzae (NTHI) is the most common bacterial pathogen associated with sinusitis and serves as proof of principle pathogen for identifying biomarkers. Study Design In vitro and in vivo studies using proteomic-based analysis of cultures of NTHI and a novel, experimental chinchilla polymicrobial sinusitis model. Methods Nano-liquid chromatography /tandem mass spectrometry (nano-LC-MS/MS) was performed to annotate the secretome from an NTHI biofilm. A model of NTHI-induced sinusitis was developed in a chinchilla, and NTHI proteins were detected in chinchilla secretions. A reference standard RT-PCR-based assay was adapted to allow for sensitivity and specificity testing of the identified signature biomarkers in human patients. Results Outer membrane proteins P2 (OMP-P2) and P5 (OMP-P5) were identified as promising candidates for the detection of NTHI biofilms and positively detected in nasopharyngeal secretions of chinchillas experimentally infected with NTHI. An RT-PCR based test for the presence of NTHI biofilms demonstrated 100% sensitivity and 100% specificity when tested against eight unique strains commonly found in human bacterial rhinosinusitis. Conclusions Proteomic analysis was successful in identifying signature proteins for possible use as a biomarker for chronic rhinosinusitis (CRS). OMP-P2 and OMP-P5 were validated as promising candidates and were positively detected from nasopharyngeal secretions from chinchillas experimentally infected with NTHI. Collectively, these data support the use of OMP-P2 and OMP-P5 as biomarkers for a human clinical trial to develop a point-of-care medical diagnostic test to assist in the diagnosis and treatment of CRS.
机译：目的/假设假设是可以通过基于蛋白质组学的高通量技术，在鼻窦分泌物中鉴定出特征性细菌蛋白。不可归类的流感嗜血杆菌（NTHI）是与鼻窦炎相关的最常见细菌病原体，可作为鉴定生物标志物的主要病原体的证明。研究设计使用基于蛋白质组学的NTHI培养物分析和新型实验性龙猫多菌性鼻窦炎模型进行体外和体内研究。方法采用纳米液相色谱/串联质谱（nano-LC-MS / MS）对NTHI生物膜的分泌蛋白进行注释。在黄鼠体内建立了NTHI诱发的鼻窦炎模型，在黄鼠分泌物中检测到NTHI蛋白。参照标准基于RT-PCR的检测方法适用于对人类患者中已鉴定的生物标志物进行敏感性和特异性测试。结果外膜蛋白P2（OMP-P2）和P5（OMP-P5）被鉴定为检测NTHI生物膜的有希望的候选物，并在实验性感染NTHI的龙猫的鼻咽分泌物中被阳性检测。当针对人类细菌性鼻-鼻窦炎中常见的八种独特菌株进行测试时，针对NTHI生物膜存在的基于RT-PCR的测试显示出100％的敏感性和100％的特异性。结论蛋白质组学分析成功地鉴定了可能用作慢性鼻-鼻窦炎（CRS）生物标志物的特征蛋白。 OMP-P2和OMP-P5被确认为有前途的候选物，并从实验性感染NTHI的龙猫的鼻咽分泌物中被阳性检测到。总体而言，这些数据支持将OMP-P2和OMP-P5用作人类临床试验的生物标志物，以开发即时医疗诊断测试以辅助CRS的诊断和治疗。

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