Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis

IF 2.1 4区医学 Q3 INFECTIOUS DISEASES

Diagnostic microbiology and infectious disease Pub Date : 2024-08-28 DOI:10.1016/j.diagmicrobio.2024.116517

Macarena Sarli , María N. De Salvo , Paula M. Díaz Pérez , Gabriel L. Cicuttin , Santiago Nava , Patrick S. Sebastian

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Abstract

The aim of this work was to develop a real-time PCR assay with a TaqMan® probe that detects a species-specific part of the 16S rDNA gene of Ehrlichia canis. Canine blood samples (n = 207), collected and tested by a conventional PCR assay within a study conducted by De Salvo et al., were simultaneously analyzed with the novel designed real-time PCR, and the results of both assays were compared. The agreement between the two methods was 97.6 % with a kappa value of 0.92186. Hereby, the standard error was 0.034416 and the 95 % confidence interval from 0.8544 to 0.98931. While the conventional PCR assay showed false negative results (2.42 %; 5/207), the real-time PCR assays showed a specificity of 100 %. The results of the current study showed that the developed assay presents sensitivity and specificity for the detection of E. canis in blood samples, adding a new tool for the diagnosis of this pathogen.

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开发和评估用于检测犬科埃立克次氏体物种特异性的 TaqMan® 实时 PCR 分析法

这项研究的目的是开发一种使用 TaqMan® 探针的实时 PCR 检测方法，该探针可检测犬艾氏原虫 16S rDNA 基因的物种特异性部分。在 De Salvo 等人进行的一项研究中，用传统 PCR 方法采集并检测了犬血样本（n = 207），同时用新设计的实时 PCR 方法对其进行了分析，并对两种方法的结果进行了比较。两种方法的一致性为 97.6%，卡帕值为 0.92186。因此，标准误差为 0.034416，95% 置信区间为 0.8544 至 0.98931。传统 PCR 检测结果显示出假阴性结果（2.42%；5/207），而实时 PCR 检测的特异性为 100%。目前的研究结果表明，所开发的检测方法对检测血液样本中的犬大肠杆菌具有灵敏性和特异性，为诊断这种病原体增添了一种新的工具。

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来源期刊

Diagnostic microbiology and infectious disease 医学-传染病学

CiteScore

5.30

自引率

3.40%

发文量

149

审稿时长

56 days

期刊介绍： Diagnostic Microbiology and Infectious Disease keeps you informed of the latest developments in clinical microbiology and the diagnosis and treatment of infectious diseases. Packed with rigorously peer-reviewed articles and studies in bacteriology, immunology, immunoserology, infectious diseases, mycology, parasitology, and virology, the journal examines new procedures, unusual cases, controversial issues, and important new literature. Diagnostic Microbiology and Infectious Disease distinguished independent editorial board, consisting of experts from many medical specialties, ensures you extensive and authoritative coverage.