Development and validation of a real-time PCR assay, and phylogenetic analysis of Anaplasma platys

IF 2 2区农林科学 Q2 PARASITOLOGY

Veterinary parasitology Pub Date : 2025-04-19 DOI:10.1016/j.vetpar.2025.110475

Nokuzola F. Nkosi , Charles Byaruhanga , Sintayehu M. Arega , Anne Conan , Darryn L. Knobel , Marinda C. Oosthuizen , Melvyn Quan

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引用次数: 0

Abstract

Anaplasmosis is a tick-borne disease caused by species of the genus Anaplasma. In dogs anaplasmosis is caused by Anaplasma phagocytophilum and Anaplasma platys. These bacteria are in the family Anaplasmataceae in the order Rickettsiales. Anaplasma platys is a Gram-negative bacterium that is of public health and veterinary importance. This pathogen exclusively infects platelets and causes infectious cyclic thrombocytopenia in dogs. Infection occurs through the bite of an infected ixodid tick, Rhipicephalus sanguineus, which is the principal vector and is also known to transmit Ehrlichia canis, another bacteria of veterinary importance. Our group recently reported on the developed group-specific Ehrlichia/Anaplasma primers and the A. platys-specific TaqMan® Minor Groove Binder probe for multiplexing purposes. This study validated the A. platys TaqMan® PCR assay targeting the 16S rRNA gene. Furthermore, phylogenetic analysis was used to characterize A. platys. The assay efficiency was 94.9 %, and the 95 % limit of detection (LOD) was 5.08 A. platys plasmid copies/μl blood with a 95 % confidence interval of 3.1–10.2. The assay did not cross-react when tested against other haemoparasites. The phylogenetic characterization of the Mnisi community samples revealed that the A. platys sequences from this area grouped with other A. platys sequences from South Africa and other countries, including India, Zambia, Taiwan, Thailand, and Croatia. The developed TaqMan® qPCR assay will be a valuable tool for the early diagnosis of A. platys by preventing inappropriate use of antibiotics and alleviating potential emerging antimicrobial resistance. Additionally, early detection and administration of the correct antibiotics speed recovery time.

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platys无原体实时PCR检测方法的建立和验证及系统发育分析

无形体病是由无形体属的一种引起的蜱传疾病。狗无形体病是由嗜吞噬细胞无形体和扁平无形体引起的。这些细菌属于无原体科立克次体目。platys无原体是一种革兰氏阴性细菌，对公共卫生和兽医具有重要意义。这种病原体专门感染血小板并引起犬的感染性循环血小板减少症。感染是通过受感染的蜱血根头蜱的叮咬发生的，蜱血根头蜱是主要的病媒，而且已知还传播另一种具有兽医重要性的细菌——犬埃利希体。我们的团队最近报道了开发的群体特异性埃利希菌/无原体引物和A. platyss特异性TaqMan®Minor Groove Binder探针，用于多路复用目的。本研究验证了针对16S rRNA基因的A. platys TaqMan®PCR检测。此外，还利用系统发育分析方法对沙蚕进行了鉴定。检测效率为94.9 %，检出限（LOD）为95 %，检出限为5.08 A。质粒拷贝数/μl血95 %置信区间为3.1 ~ 10.2。当与其他血液寄生虫试验时，该试验没有交叉反应。通过对Mnisi群落样本的系统发育分析，发现该地区的platys序列与其他地区的platys序列具有类群性。TaqMan®qPCR检测将为platys的早期诊断提供有价值的工具，可防止不适当使用抗生素并减轻潜在的抗菌素耐药性。此外，早期发现和使用正确的抗生素可以加快恢复时间。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Veterinary parasitology 农林科学-寄生虫学

CiteScore

5.30

自引率

7.70%

发文量

126

审稿时长

36 days

期刊介绍： The journal Veterinary Parasitology has an open access mirror journal,Veterinary Parasitology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. This journal is concerned with those aspects of helminthology, protozoology and entomology which are of interest to animal health investigators, veterinary practitioners and others with a special interest in parasitology. Papers of the highest quality dealing with all aspects of disease prevention, pathology, treatment, epidemiology, and control of parasites in all domesticated animals, fall within the scope of the journal. Papers of geographically limited (local) interest which are not of interest to an international audience will not be accepted. Authors who submit papers based on local data will need to indicate why their paper is relevant to a broader readership. Parasitological studies on laboratory animals fall within the scope of the journal only if they provide a reasonably close model of a disease of domestic animals. Additionally the journal will consider papers relating to wildlife species where they may act as disease reservoirs to domestic animals, or as a zoonotic reservoir. Case studies considered to be unique or of specific interest to the journal, will also be considered on occasions at the Editors'' discretion. Papers dealing exclusively with the taxonomy of parasites do not fall within the scope of the journal.