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利用嵌合蛋白 rGroEL-FLAG300 作为抗原的间接 ELISA 增强对类鼻疽病的血清学诊断。

Enhanced serodiagnosis of melioidosis by indirect ELISA using the chimeric protein rGroEL-FLAG300 as an antigen.

机构信息

Department of Basic Medical Science, Faculty of Medicine, Vajira Hospital, Navamindradhiraj University, Bangkok, 10300, Thailand.

Mahidol Oxford Tropical Medicine Research Unit, Bangkok, 10400, Thailand.

出版信息

BMC Infect Dis. 2022 Apr 19;22(1):387. doi: 10.1186/s12879-022-07369-4.

Abstract

BACKGROUND

The accurate and rapid diagnosis of melioidosis is challenging. Several serological approaches have been developed using recombinant antigens to improve the diagnostic indices of serological tests for melioidosis.

METHODS

Fusion proteins from Burkholderia pseudomallei (rGroEL-FLAG300) were evaluated as a potential target antigen for melioidosis antibodies. A total of 220 serum samples from 38 culture proven melioidosis patients (gold standard), 126 healthy individuals from endemic (n = 37) and non-endemic (n = 89) Thai provinces and 56 patients with other proven bacterial infections as negative controls were tested using indirect enzyme-linked immunosorbent assays (ELISA).

RESULTS

Using an optical density (OD) cut-off of 0.299148, our assay had 94.74% sensitivity (95% confidence interval (CI) = 82.3-99.4%), 95.05% specificity (95% CI = 90.8-97.7%), and 95% accuracy, which was better than in our previous work (90.48% sensitivity, 87.14% specificity, and 87.63% accuracy).

CONCLUSION

Our results suggest that the application of chimeric antigens in ELISA could improve the serological diagnosis of melioidosis and should be reconfirmed with greater patient numbers.

摘要

背景

准确快速诊断类鼻疽具有挑战性。已经开发了几种使用重组抗原的血清学方法,以提高类鼻疽血清学检测的诊断指标。

方法

使用伯氏考克斯体(rGroEL-FLAG300)融合蛋白作为类鼻疽抗体的潜在靶抗原进行评估。共检测了 220 份血清样本,其中 38 份来自经培养证实的类鼻疽患者(金标准),126 份来自地方性(n=37)和非地方性(n=89)泰国省份的健康个体,56 份来自其他已证实的细菌性感染患者作为阴性对照,使用间接酶联免疫吸附试验(ELISA)进行检测。

结果

使用 0.299148 的光密度(OD)截断值,我们的检测方法的敏感性为 94.74%(95%置信区间(CI)=82.3-99.4%),特异性为 95.05%(95%CI=90.8-97.7%),准确率为 95%,优于我们之前的工作(敏感性 90.48%,特异性 87.14%,准确率 87.63%)。

结论

我们的结果表明,嵌合抗原在 ELISA 中的应用可以提高类鼻疽的血清学诊断,应该用更多的患者数量进行重新确认。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43c7/9020111/e6ff71fb990d/12879_2022_7369_Fig1_HTML.jpg

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