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下一代 DNA 测序在诊断方面优于传统的培养检测。

Next-generation DNA sequencing offers diagnostic advantages over traditional culture testing.

机构信息

Zymo Research Corporation, Irvine, CA.

Long Island Bird and Exotics Veterinary Clinic, Great Neck, NY.

出版信息

Am J Vet Res. 2023 Jun 12;84(8). doi: 10.2460/ajvr.23.03.0054. Print 2023 Aug 1.

Abstract

OBJECTIVE

While the clinical utility of next-generation DNA sequencing (NGS) as a diagnostic tool for infections in humans and traditional pets has been demonstrated, there is a lack of data regarding its utility for exotic animals. For exotic patients, traditional culturing is especially challenging for anaerobic and fungal pathogens. Therefore, diagnosis often relies on PCR, which provides a high degree of sensitivity and specificity, although it targets only a predetermined, finite pathogen panel. NGS provides the same benefits as PCR, while also offering de novo identification and quantification of all bacteria and fungi present in a clinical sample, including novel pathogen discovery.

PROCEDURES

Clinical samples from 78 exotic animal patients were collected simultaneously for conventional culture testing and NGS analysis. Results provided by each laboratory were compared for the presence and absence of bacterial and fungal pathogens and commensals.

RESULTS

Results showed large bacterial and fungal species diversity in the study cohort and a lack of sensitivity of microbial culture testing. Culture failed to grow 15% of putative bacterial and 81% of putative fungal pathogens that were identified by NGS. The probability of a "no growth" diagnosis was 14% higher for bacteria and 49% higher for fungi with culture versus NGS testing if fungal culture was conducted.

CLINICAL RELEVANCE

Culture testing failed to diagnose a substantial number of both bacterial and fungal pathogens, which were detected by NGS. This highlights the limitations of traditional culture-based testing and displays the clinically advanced utility of NGS-based diagnostics in exotic animal medicine.

摘要

目的

虽然下一代 DNA 测序(NGS)作为人类和传统宠物感染的诊断工具的临床实用性已经得到证实,但关于其在异宠中的应用的数据还很缺乏。对于异宠患者来说,传统的培养方法对于厌氧菌和真菌病原体尤其具有挑战性。因此,诊断通常依赖于 PCR,PCR 具有高度的灵敏度和特异性,尽管它只针对预定的、有限的病原体面板。NGS 提供了与 PCR 相同的好处,同时还可以对临床样本中存在的所有细菌和真菌进行从头鉴定和定量,包括新病原体的发现。

方法

同时从 78 只异宠患者身上采集临床样本,用于传统培养检测和 NGS 分析。比较每个实验室提供的结果,以确定是否存在细菌和真菌病原体及共生菌。

结果

结果表明,研究队列中存在大量的细菌和真菌物种多样性,微生物培养检测的敏感性较低。培养未能培养出 NGS 鉴定出的 15%的假定细菌和 81%的假定真菌病原体。如果进行真菌培养,与 NGS 检测相比,培养法诊断“无生长”的概率分别高出细菌 14%,真菌 49%。

临床相关性

培养检测未能诊断出大量的细菌和真菌病原体,这些病原体被 NGS 检测到。这突显了传统基于培养的检测方法的局限性,并展示了 NGS 为基础的诊断方法在异宠医学中的临床先进应用。

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