Department of Veterinary Parasitology, College of Veterinary Science, Guru Angad Dev Veterinary & Animal Sciences University, Ludhiana,, Punjab, India.
Department of Dairy Microbiology, College of Dairy & Food Science Technology, Guru Angad Dev Veterinary & Animal Sciences University, Ludhiana,, Punjab, India.
Acta Parasitol. 2024 Sep;69(3):1458-1470. doi: 10.1007/s11686-024-00856-5. Epub 2024 Aug 16.
Tick-transmitted parasites as Babesia gibsoni, Babesia vogeli, Ehrlichia canis, and Hepatozoon canis are major health concern for dogs. Owing to prevalence and infection severity, there is need of sensitive, specific, and affordable test for their simultaneous detection.
Prevalence of B. gibsoni, B. vogeli, E. canis, and H. canis infections was assessed on 719 blood samples by microscopy and multiplex PCR assay targeting 18S rRNA (B. gibsoni & H. canis), ITS1 & 5.8S rRNA (B. vogeli) and VirB9 gene (E. canis). An internal control (canine-actin) was also included to increase the accuracy of assay and effect of associated risk factors with disease prevalence was also studied.
Microscopic prevalence of B. gibsoni, B. vogeli, E. canis and H. canis was 5.0%, 0.1%, 1.4% and 1.0%, respectively, whereas with multiplex PCR assay, the corresponding values were 8.9%, 1.1%, 2.6% and 5.1% besides concurrent infections of B. gibsoni & H. canis (0.4%), B. gibsoni & E. canis (0.4%), E. canis & H. canis (0.3%) and B. gibsoni & B. vogeli (0.1%). Analytical sensitivity of developed assay was 0.1pg (B. gibsoni & H. canis), 0.01pg (B. vogeli), and 1.0pg (E. canis). A ″fair″ (B. vogeli & H. canis) to ″substantial″ (B. gibsoni & E. canis) agreement between two tests was observed with data as statistically significant. Breed, sex and location were significantly associated with B. gibsoni infection.
The developed multiplex PCR assay offers a potential solution to detect these pathogens simultaneously, aiding in timely diagnosis and effective disease management in suspected dogs.
蜱传播的寄生虫，如巴贝斯虫、巴贝斯虫、犬埃立克体和犬肝孢子虫，是犬类健康的主要关注点。由于其流行程度和感染严重程度，需要一种敏感、特异和经济实惠的方法来同时检测这些寄生虫。
通过显微镜检查和针对 18S rRNA（巴贝斯虫和犬肝孢子虫）、ITS1 和 5.8S rRNA（巴贝斯虫）和 VirB9 基因（犬埃立克体）的多重 PCR 检测，评估了 719 份血液样本中巴贝斯虫、巴贝斯虫、犬埃立克体和犬肝孢子虫感染的流行率。还包括了一个内参（犬肌动蛋白），以提高检测的准确性，并研究与疾病流行率相关的风险因素的影响。
巴贝斯虫、巴贝斯虫、犬埃立克体和犬肝孢子虫的显微镜检测流行率分别为 5.0%、0.1%、1.4%和 1.0%，而多重 PCR 检测的相应值分别为 8.9%、1.1%、2.6%和 5.1%，此外还存在巴贝斯虫和犬肝孢子虫（0.4%）、巴贝斯虫和犬埃立克体（0.4%）、犬埃立克体和犬肝孢子虫（0.3%）以及巴贝斯虫和巴贝斯虫（0.1%）的并发感染。该检测方法的分析灵敏度为 0.1pg（巴贝斯虫和犬肝孢子虫）、0.01pg（巴贝斯虫）和 1.0pg（犬埃立克体）。两种检测方法之间的一致性为“良好”（巴贝斯虫和犬肝孢子虫）到“中等”（巴贝斯虫和犬埃立克体），数据具有统计学意义。品种、性别和地理位置与巴贝斯虫感染显著相关。
该开发的多重 PCR 检测方法为同时检测这些病原体提供了一种潜在的解决方案，有助于在疑似犬只中及时诊断和有效管理疾病。
