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使用贝叶斯潜在类别分析比较传统聚合酶链反应(PCR)和两种便于现场操作的检测方法在蜱虫中检测DNA的检测性能。

Comparison of test performance of a conventional PCR and two field-friendly tests to detect DNA in ticks using Bayesian latent class analysis.

作者信息

Kamau Maureen W, Witte Carmel, Goosen Wynand, Mutinda Mathew, Villinger Jandouwe, Getange Dennis, Khogali Rua, von Fricken Michael E, Fèvre Eric Maurice, Zimmerman Dawn, Linton Yvonne-Marie, Miller Michele

机构信息

Mpala Research Centre, Nanyuki, Kenya.

Division of Molecular Biology and Human Genetics, Department of Science, and Innovation - National Research Foundation Centre of Excellence for Biomedical Tuberculosis Research, Faculty of Medicine and Health Sciences, South African Medical Research Council Centre for Tuberculosis Research, Stellenbosch University, Stellenbosch, South Africa.

出版信息

Front Vet Sci. 2024 Jun 19;11:1396714. doi: 10.3389/fvets.2024.1396714. eCollection 2024.

Abstract

INTRODUCTION

()-infected livestock and wildlife have been epidemiologically linked to human Q fever outbreaks. Despite this growing zoonotic threat, knowledge of coxiellosis in wild animals remains limited, and studies to understand their epidemiologic role are needed. In -endemic areas, ticks have been reported to harbor and spread and may serve as indicators of risk of infection in wild animal habitats. Therefore, the aim of this study was to compare molecular techniques for detecting DNA in ticks.

METHODS

In total, 169 ticks from wild animals and cattle in wildlife conservancies in northern Kenya were screened for DNA using a conventional PCR (cPCR) and two field-friendly techniques: Biomeme's qPCR Go-strips (Biomeme) and a new PCR high-resolution melt (PCR-HRM) analysis assay. Results were evaluated, in the absence of a gold standard test, using Bayesian latent class analysis (BLCA) to characterize the proportion of positive ticks and estimate sensitivity (Se) and specificity (Sp) of the three tests.

RESULTS

The final BLCA model included main effects and estimated that PCR-HRM had the highest Se (86%; 95% credible interval: 56-99%), followed by the Biomeme (Se = 57%; 95% credible interval: 34-90%), with the estimated Se of the cPCR being the lowest (24%, 95% credible interval: 10-47%). Specificity estimates for all three assays ranged from 94 to 98%. Based on the model, an estimated 16% of ticks had DNA present.

DISCUSSION

These results reflect the endemicity of in northern Kenya and show the promise of the PCR-HRM assay for surveillance in ticks. Further studies using ticks and wild animal samples will enhance understanding of the epidemiological role of ticks in Q fever.

摘要

引言

感染了()的家畜和野生动物在流行病学上与人类Q热疫情有关。尽管这种人畜共患病威胁日益增加,但关于野生动物柯克斯体病的知识仍然有限,需要开展研究以了解它们在流行病学中的作用。在()流行地区,据报道蜱虫携带并传播(),可能是野生动物栖息地感染风险的指标。因此,本研究的目的是比较检测蜱虫中()DNA的分子技术。

方法

使用常规PCR(cPCR)以及两种便于现场操作的技术:Biomeme的()qPCR Go试纸条(Biomeme)和一种新的()PCR高分辨率熔解(PCR-HRM)分析方法,对肯尼亚北部野生动物保护区内来自野生动物和牛的总共169只蜱虫进行()DNA筛查。在没有金标准检测方法的情况下,使用贝叶斯潜在类别分析(BLCA)评估结果,以确定()阳性蜱虫的比例,并估计这三种检测方法的灵敏度(Se)和特异性(Sp)。

结果

最终的BLCA模型纳入了主要效应,估计PCR-HRM的Se最高(86%;95%可信区间:56-99%),其次是Biomeme(Se = 57%;95%可信区间:34-90%),cPCR的估计Se最低(24%,95%可信区间:10-47%)。所有三种检测方法的特异性估计范围为94%至98%。根据该模型,估计有16%的蜱虫存在()DNA。

讨论

这些结果反映了肯尼亚北部()的地方性流行情况,并显示了PCR-HRM检测方法在蜱虫监测方面的前景。使用蜱虫和野生动物样本的进一步研究将增进对蜱虫在Q热流行病学中作用的理解。

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