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用于检测人血清中球孢子菌几丁质酶-1(CTS1)的定量抗原检测方法的开发。

Development of a Quantitative Antigen Assay to Detect Coccidioidal Chitinase-1 (CTS1) in Human Serum.

作者信息

Grill Francisca J, Grys Thomas E, Grill Marie F, Roeder Alexa, Blair Janis E, Lake Douglas F

机构信息

School of Life Sciences, Arizona State University, Tempe, Arizona, USA.

Department of Laboratory Medicine and Pathology, Mayo Clinic, Phoenix, Arizona, USA.

出版信息

Open Forum Infect Dis. 2021 Jun 28;8(7):ofab344. doi: 10.1093/ofid/ofab344. eCollection 2021 Jul.

Abstract

BACKGROUND

Coccidioidomycosis is often diagnosed with a collection of tests that rely on the patient's ability to mount an immune response to the fungus (antibody-based diagnostics), making diagnosis of this infection challenging. Here we present an antigen-based assay that detects and quantifies coccidioidal chitinase-1 (CTS1) in human serum.

METHODS

An inhibition-based enzyme-linked immunoassay (ELISA) was developed that utilizes a monoclonal antibody specific for coccidioidal CTS1. CTS1 was quantified in commercial antigen preparations using recombinant CTS1 as a standard. Sera from 192 individuals from an endemic area were tested, which included 78 patients (40.6%) with proven or probable coccidioidomycosis.

RESULTS

The quantity of CTS1 in diagnostic commercial antigen preparations from different suppliers varied. CTS1 antigenemia was detected in 87.2% of patients with proven or probable coccidioidomycosis. Specificity was determined to be 96.94% using serum from individuals who reside in the Phoenix, Arizona area who did not have coccidioidomycosis. Levels of CTS1 correlated with low- and high-titer serology from patients with a coccidioidomycosis diagnosis.

CONCLUSIONS

Since the CTS1 inhibition ELISA described in this report does not depend on the host immune response, it is a promising diagnostic tool to aid in diagnosis and disease monitoring of coccidioidomycosis.

摘要

背景

球孢子菌病通常通过一系列依赖患者对该真菌产生免疫反应能力的检测方法来诊断(基于抗体的诊断方法),这使得这种感染的诊断具有挑战性。在此,我们介绍一种基于抗原的检测方法,可检测和定量人血清中的球孢子菌几丁质酶-1(CTS1)。

方法

开发了一种基于抑制的酶联免疫吸附测定(ELISA),该方法利用对球孢子菌CTS1具有特异性的单克隆抗体。使用重组CTS1作为标准品,对商业抗原制剂中的CTS1进行定量。对来自流行地区的192名个体的血清进行了检测,其中包括78例确诊或疑似球孢子菌病的患者(40.6%)。

结果

不同供应商的诊断用商业抗原制剂中CTS1的含量有所不同。在87.2%的确诊或疑似球孢子菌病患者中检测到CTS1抗原血症。使用来自亚利桑那州凤凰城地区无球孢子菌病的个体的血清,确定特异性为96.94%。CTS1水平与球孢子菌病诊断患者的低滴度和高滴度血清学结果相关。

结论

由于本报告中描述的CTS1抑制ELISA不依赖于宿主免疫反应,因此它是一种有前景的诊断工具,有助于球孢子菌病的诊断和疾病监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/286c/8320284/74fd48db1ec6/ofab344f1.jpg

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