Simultaneous differential detection of canine blood parasites: Multiplex high-resolution melting analysis (mHRM)

Buddhachat, Kittisak; Meerod, Tirawit; Pradit, Waranee; Siengdee, Puntita; Chomdej, Siriwadee; Nganvongpanit, Korakot

doi:10.1016/j.ttbdis.2020.101370

Cited by 16 publications

(17 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The most PCR-detected tick-transmitted bacterium contracted by canines was E. canis (38.3%), the causative agent of canine monocytic ehrlichiosis. In agreement with this study, E. canis was the most prevalent tick-borne bacterium (21.5-36%) reported in northern Thailand [17,20,21]. In other Southeast Asian countries, the relatively high occurrence of E. canis is consistent with previous studies reporting 21.8%, 15.7%, 25.8%, 5.3%, 11.1%, and 36.2% in Cambodia [7], the Philippines [3], Vietnam, Singapore, Malaysia, and Indonesia [22], respectively.…”

Section: Discussionsupporting

confidence: 91%

“…The documented rates of CTBPs in dogs varied in several epidemiological surveys conducted in different geographical areas in Thailand [ 11 , 17 , 18 , 19 ]; however, only a few studies of Vector-borne pathogens have been done based on vector. Our results indicated that CTBPs including A. platys , B. vogeli , E. canis , M. haemocanis and H. canis are endemic in the studied area.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular Detection of Tick-Borne Pathogens in Stray Dogs and Rhipicephalus sanguineus sensu lato Ticks from Bangkok, Thailand

Phoosangwalthong

Kamyingkird

et al. 2021

Pathogens

View full text Add to dashboard Cite

Canine tick-borne pathogens (CTBPs) such as Babesia vogeli, Ehrlichia canis, Anaplasma platys, Hepatozoon canis, and Mycoplasma haemocanis are important pathogens in dogs worldwide. Rhipicephalus sanguineus sensu lato, the main vector of several CTBPs, is the most common tick species found on dogs in Thailand. The present study identified CTBPs in dogs and ticks infested dogs. Samples (360 dog blood samples and 85 individual ticks) were collected from stray dogs residing in 37 temples from 24 districts in Bangkok and screened for CTBPs using molecular techniques. The most common CTBP found infecting dogs in this study was Ehrlichia canis (38.3%) followed by Mycoplasma haemocanis (34.2%), Hepatozoon canis (19.7%), Babesia vogeli (18.1%), and Anaplasma platys (13.9%), respectively. Furthermore, A. platys (22.4%) was the most common CTBP in ticks followed by M. haemocanis (18.8%), B. vogeli (9.4%), H. canis (5.9%), and E. canis (2.4%), respectively. The detection of CTBPs from the present study highlights the potential risk of infections that may occur in stray dogs and their ticks residing in Bangkok temples. These findings underline the importance of performing active surveys to understand the complexity of distributions of CTBPs in dogs and their ticks in Thailand.

show abstract

Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Molecular Detection of Tick-Borne Pathogens in Stray Dogs and Rhipicephalus sanguineus sensu lato Ticks from Bangkok, Thailand

Phoosangwalthong

Kamyingkird

et al. 2021

Pathogens

View full text Add to dashboard Cite

show abstract

“…The occurrence of CVBPs were frequent in stray dogs in studied areas (43.1%, 75/174), of which H. canis , the causative agent of canine hepatozoonosis, was the most PCR-detected intracellular protozoan parasite (24.7%, 43/174) contracted by canine. The detection rate of these pathogens varies compared to previous reports conducted in the northern areas (41.2%, 28/68) [ 21 ] and in the southern areas (43.1%, 78/181) [ 22 ]. The variability observed in this study with regard to others could be due to the difference in the number of dogs, the selection criteria, the sampling size, the geographical area, and the sampling season [ 6 , 22 ].…”

Section: Discussionmentioning

confidence: 72%

First Molecular Detection of Babesia gibsoni in Stray Dogs from Thailand

Ngasaman

Saechan

et al. 2021

Pathogens

View full text Add to dashboard Cite

In southern Thailand, the increasingly growing population of stray dogs is a concern to public health and environmental safety because of the lack of medical attention and control. More importantly, these animals are considered reservoirs for many zoonotic pathogens. The objective of this study was to molecularly detect canine vector-borne pathogens, and to perform genetic characterization of Babesia gibsoni present in stray dogs from southern Thailand. Blood samples were collected from 174 stray dogs in two provinces (Songkhla and Narathiwat) in southern Thailand. PCR analyses were executed using specific primers based on the Babesia spp. 18S rRNA gene, Babesia gibsoni Internal transcribed spacer 1 (ITS1) region, Ehrlichia canis citrate synthase (gltA) gene, Hepatozoon spp. 18S rRNA gene and Anaplasma platys heat shock protein (groEL) gene. The most common canine vector-borne pathogen found infecting stray dogs in this study was Hepatozoon canis (24.7%) followed by A. platys (14.9%), Babesia vogeli (8.0%), B. gibsoni (6.3%), and E. canis (1.72%). Concurrent infection with more than one pathogen occurred in 72 cases. Phylogenetic analysis based on the ITS1 region and 18S rRNA gene revealed that the B. gibsoni isolates from this study shared a large proportion of their identities with each other and with other reported B. gibsoni genotypes from Asia. This study highlights the molecular detection of B. gibsoni in dogs in Thailand for the first time and presents the genetic characterization by sequencing the ITS1 region and 18S rRNA gene of B. gibsoni from Thailand. Follow-up studies are needed to elucidate the origin, distribution, and vectors of B. gibsoni parasites circulating in dogs in Thailand, as well as to determine to what extent dogs are important reservoir hosts for zoonotic canine vector-borne disease infection in the studied area.

show abstract

“…Real-time PCR assays for the detection of A. platys, E. canis, haemotropic Mycoplasma spp., B. vogeli, B. gibsoni and H. canis have been developed previously, although none of these methods have targeted all these VBP agents simultaneously and only a proportion have utilised highly specific, Taq-Man probe-based chemistry [9,[24][25][26]28,32,38,47,[49][50][51][52][53][54][55].…”

Section: Discussionmentioning

confidence: 99%

“…Given the breadth of potential canine VBPs, accurate diagnosis of the responsible species is critically important as each may require different treatment protocols to cure the infection [8,31]. Diagnosis has historically been conducted by microscopic examination of stained blood smears, a method notorious for its poor sensitivity, that also requires substantial expertise on the part of the microscopist to distinguish morphologically similar pathogens [9,32,33]. Some of these issues are also encountered when conducting serological diagnosis, e.g., low sensitivity or poor specificity due to antibody cross-reactivity [30].…”

Section: Introductionmentioning

confidence: 99%

Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific

et al. 2021

View full text Add to dashboard Cite

The Asia-Pacific hosts a large diversity of canine vector-borne pathogens (VBPs) with some of the most common and most pathogenic, generating significant mortality as well as a spectrum of health impacts on local dog populations. The VBPs Anaplasma platys, Babesia gibsoni, Babesia vogeli, Ehrlichia canis, Hepatozoon canis and haemotropic Mycoplasma spp. are all endemic throughout the region, with many exhibiting shifting geographical distributions that warrant urgent attention. Moreover, many of these species cause similar clinical signs when parasitising canine hosts, whilst knowledge of the exact pathogen is critical to ensure treatment is effective. This is complicated by frequent coinfection that can exacerbate pathology. Here, we describe the development, optimisation and validation of two novel quadruplex Taq-Man based real-time PCRs (qPCRs) for the specific and sensitive detection of the aforementioned VBPs. To ensure accurate evaluation of diagnostic performance, results of our qPCRs were evaluated on field samples from Thai dogs and compared with both conventional PCR (cPCR) results and next-generation sequencing (NGS) metabarcoding. Our qPCRs were found to be more sensitive at detecting canine VBP than cPCR and generated results similar to those achieved by NGS. These qPCRs will provide a valuable high-throughput diagnostic tool available to epidemiologists, researchers and clinicians for the diagnosis of key canine VBPs in the Asia-Pacific and further afield.

show abstract

Simultaneous differential detection of canine blood parasites: Multiplex high-resolution melting analysis (mHRM)

Cited by 16 publications

References 36 publications

Molecular Detection of Tick-Borne Pathogens in Stray Dogs and Rhipicephalus sanguineus sensu lato Ticks from Bangkok, Thailand

Molecular Detection of Tick-Borne Pathogens in Stray Dogs and Rhipicephalus sanguineus sensu lato Ticks from Bangkok, Thailand

First Molecular Detection of Babesia gibsoni in Stray Dogs from Thailand

Novel High-Throughput Multiplex qPCRs for the Detection of Canine Vector-Borne Pathogens in the Asia-Pacific

Contact Info

Product

Resources

About