Canine babesiosis is a serious infectious disease in subtropical and tropical regions. Typically, clinical detection of canine babesiosis is performed by blood smear observation or the traditional polymerase chain reaction (PCR). Herein, we developed a new TaqMan-based real-time PCR assay combined with a customized portable real-time PCR platform for a rapid and accurate detection of canine babesiosis. Two new primer/probe pairs (B18S and BITS1) were designed based on 18S ribosomal RNA and an internal transcribed spacer 1 (ITS1) sequence to differentiate Babesia canis and B. gibsoni (Asian genotype) DNAs from canine blood samples. Additionally, a corresponding customized compact real-time PCR platform with low 6-carboxyfluorescein fluorescence detection (≤5 nM), including a fast and accurate thermal cycling ability with a user-friendly interface for thermal control and data analysis, was designed for the limited space use. Both assays (B18S and BITS1) demonstrated a sensitivity of 100 copies/reaction based on the 95 % confidence interval evaluation method. The self-developed customized portable real-time PCR analyzer presented high repeatability and reproducibility with the TaqMan-based assay. Moreover, 501 clinical specimens were collected for evaluating the performance of the proposed PCR. The positive and negative predictive values were 90 % (18 of 20) and 100 % (226 of 226), respectively, for samples suspected with B. canis infection and 98 % (55 of 56) and 100 % (199 of 199), respectively, for samples suspected with B. gibsoni infection.

犬幼犬病在亚热带和热带地区是一种严重的传染病。通常，通过血液涂片观察或传统的聚合酶链反应（PCR）进行犬幼虫病的临床检测。本文中，我们开发了一种新的基于TaqMan的实时PCR分析方法，并结合了定制的便携式实时PCR平台，可以快速，准确地检测犬幼犬病。基于18S核糖体RNA和内部转录间隔区1（ITS1）序列设计了两个新的引物/探针对（B18S和BITS1），以区分犬贝贝斯病菌和吉布斯犬（亚洲基因型）犬血样本中的DNA。此外，针对有限的用户设计了相应的定制化紧凑型实时PCR平台，该平台具有低6-羧基荧光素荧光检测（≤5nM），包括快速和准确的热循环功能以及易于使用的热控制和数据分析界面。空间使用。两种测定（B18S和BITS1）均显示出基于95％置信区间评估方法的100拷贝/反应的敏感性。自行开发的定制便携式实时PCR分析仪通过基于TaqMan的测定法具有很高的可重复性和可重复性。此外，收集了501个临床标本，以评估拟议PCR的性能。对于疑似感染者的样本，阳性和阴性预测值分别为90％（20个中的18个）和100％（226个中的226个）。B.犬感染和98％（56 55）和100％（199 199），分别用于与疑似样品B. gibsoni感染。