A novel PCR-based point-of-care method enables rapid, sensitive and reliable diagnosis of Babesia gibsoni infection in dogs

Liu, I-Li; Chi, Nai-Yu; Chang, Chia‐Ling; Hung, Ming-Long; Chiu, Chun-Ta; Chen, Hui-Wen

doi:10.1186/s12917-019-2181-5

Cited by 4 publications

(2 citation statements)

References 21 publications

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“…This method also showed Recently, the timely diagnosis of B. gibsoni by a QubeMDx PCR system was achieved with sensitivity limits between 0.000576 and 0.002% of parasitemia. The entire process was completed within 30 min, demonstrating a novel and reliable point-of-care testing method in clinical stations 36 .…”

Section: Discussionmentioning

confidence: 99%

Sensitive and rapid detection of Babesia species in dogs by recombinase polymerase amplification with lateral flow dipstick (RPA-LFD)

Onchan

Ritbamrung

Changtor

et al. 2022

Sci Rep

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Canine babesiosis is a tick-borne disease caused by Babesia spp., which infects and destroys healthy erythrocytes, leading to mortality and morbidity in dogs. The diagnosis of babesiosis is tedious and time-consuming, especially in latent and chronic infections. Here, a recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD) assay was developed for rapid and accurate detection of Babesia spp. in canine blood specimens based on the 18S rRNA region. The RPA-LFD assay using rpaBab264 gave specificity to Babesia spp. in dogs (B. vogeli and B. gibsoni) without cross-amplification to other parasites (apicomplexans and non-apicomplexans), with detection limit of at least 22.5 copies/μl (0.1 fg/µl) at 40 °C for at least 10 min. The whole process of DNA amplification by RPA and readout by LFD did not exceed 30 min. To determine the performance of the RPA-LFD assay, a total of 30 clinical samples was examined and compared with conventional PCR (cPCR) and multiplex HRM (mHRM). Eight dogs (26.67%) were detected as positive by RPA-LFD, while seven and six were found positive by cPCR and mHRM, respectively. RPA-LFD and cPCR showed high agreement with Babesia spp. detection with kappa > 0.9. We confirmed that the dogs were infected by B. vogeli from sequences of positive PCR results. Our findings suggested that RPA-LFD using the rpaBab264 assay offered a rapid, accurate, cost-effective and simple method for Babesia spp. detection that is feasibly applicable to be rapid kit at a pet hospital or point-of-care testing.

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Section: Discussionmentioning

confidence: 99%

Sensitive and rapid detection of Babesia species in dogs by recombinase polymerase amplification with lateral flow dipstick (RPA-LFD)

Onchan

Ritbamrung

Changtor

et al. 2022

Sci Rep

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“…A high κ value signifies high agreement; otherwise, it means low agreement. 20 The κ test was conducted to evaluate agreement between the serological test and the kits.…”

Section: Methodsmentioning

confidence: 99%

Performance Analysis of Novel Nucleic Acid Detection Kit for Mycoplasma pneumoniae

Wan

Zhao

et al. 2022

Clin Pediatr (Phila)

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Respiratory tract infections caused by Mycoplasma pneumoniae is a serious risk for child health. It has been difficult to prevent and control for a variety of reasons; therefore, timely diagnosis is particularly important for treatment of patients. At present, the rapid M pneumoniae test kits based on nucleic acid amplification have been commercialized and used as primary diagnostic tools for M pneumoniae infection, but current kits are time-consuming, which is difficult to meet the requirement for accurate and rapid diagnosis of M pneumoniae during epidemics. Rapid and accurate test kits are urgently required to diagnose M pneumoniae infection. In this article, we evaluated the performance of a novel nucleic acid detection kit (A) for M pneumoniae from feasibility and sensitivity, and compared it with kit B. Results showed this kit has the advantage of being rapid, sensitive, and specific, which meets the demands for the diagnosis of M pneumoniae infection in clinical settings.

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Detection of Canine Parvovirus Type 2 by a Commercially Available in-House PCR

Yen

Hoang

Wang³

et al. 2020

Taiwan Vet J

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Canine parvovirus type 2 (CPV-2) is an important pathogen that causes anorexia, vomiting, acute gastroenteritis, and bloody diarrhea. CPV-2 was divided into three variants (2a–2c) based on residue 426 of the VP2 protein. Recently, a novel Asian CPV-2c isolate was more prevalent in the Asian continent. The diagnostic tools of CPV-2 are characterized into traditional (such as immunochromatography test) and molecular methods based on their pathogen detection mechanisms. However, the low sensitivity of the immunochromatographic tests is supported by their simplicity and rapid ability to provide results for CPV-2. Surprisingly, some immunochromatography tests showed low sensitivity for the detection of novel CPV-2c compared to a real-time polymerase chain reaction (PCR) assay. Sixty rectal swabs were collected from naturally infected dogs with CPV-2 at the Animal Disease Diagnostic Center, National Pingtung University of Science and Technology, Taiwan, from 2016 to 2019. A novel in-house QubeMDx PCR system was used for diagnosis of three CPV-2 variants and compared to real-time PCR diagnosis. This diagnostic assay detected all of CPV-2 variants within 30 min and the sensitivity was identical to a real-time PCR assay using clinical samples. This study provides evidence that this novel, commercially available in-house PCR assay (QubeMDx PCR system) provides useful, simple, sensitive, reliable, and rapid diagnosis of three CPV-2 variants in veterinary clinics.

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A novel PCR-based point-of-care method enables rapid, sensitive and reliable diagnosis of Babesia gibsoni infection in dogs

Cited by 4 publications

References 21 publications

Sensitive and rapid detection of Babesia species in dogs by recombinase polymerase amplification with lateral flow dipstick (RPA-LFD)

Sensitive and rapid detection of Babesia species in dogs by recombinase polymerase amplification with lateral flow dipstick (RPA-LFD)

Performance Analysis of Novel Nucleic Acid Detection Kit for Mycoplasma pneumoniae

Detection of Canine Parvovirus Type 2 by a Commercially Available in-House PCR

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