Development and validation of a multiplex, real-time PCR assay for Babesia rossi and Babesia vogeli

Troskie, Milana; Villiers, Lourens de; Leisewitz, Andrew L.; Oosthuizen, Marinda C.; Quan, Melvyn

doi:10.1016/j.ttbdis.2018.12.004

Cited by 16 publications

(11 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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%

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

Section: Discussionmentioning

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

show abstract

“…The difference in the rate of positivity between immunofluorescence and PCR in the study population may reflect previous exposure to piroplasmids, with maintenance of antibody titers without PCR detection of parasites or because of the low sensitivity of PCR in samples collected from Babesia-infected asymptomatic dogs or dogs in the chronic phase of the disease (BOOZER; MACINTIRE, 2003). Although conventional PCR is highly sensitive and specific, the use of alternative techniques such as nested-PCR and real-time PCR (TROSKIE et al, 2018) may increase sensitivity. PCR may be particularly useful in areas endemic to Leishmania infantum, where DNA from other vector-borne pathogens, including B. vogeli, is detected in L. infantum-seropositive dogs, reinforcing the importance of molecular tests for detecting the causative agents of vector-borne diseases in these areas (SOUSA et al, 2013), including Teresina, which is endemic for canine visceral leishmaniasis (DRUMOND; COSTA, 2011).…”

Section: Discussionmentioning

confidence: 99%

Molecular, serological, and parasitological detection of Babesia vogeli in dogs in the state of Piauí, Brazil

Braga

Souza

Silva

et al. 2019

SCA

View full text Add to dashboard Cite

Studies on canine babesiosis in northeastern Brazil are scarce, although the weather conditions in this region are favorable for the development of the tick vector. This study determined the prevalence of Babesia vogeli in dogs sampled in Teresina, state of Piauí, northeast Brazil, using direct and indirect diagnostic methods and performed a phylogenetic analysis of 18S rRNA sequences. A total of 315 dogs were screened during routine care regardless of clinical suspicion. Blood was collected by jugular venipuncture to perform indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR) and for parasite screening in peripheral blood smears. Positivity was 2.2% (7/315) by microscopy, 4.8% (15/315) by PCR, and 48.6% (153/315) by IFA. PCR amplified a 602-bp fragment of the piroplasmid 18S rRNA gene, and sequence alignment and analysis revealed 99% homology with B. vogeli isolates from other regions of Brazil and other countries. In addition, there was high variability among sequences from other northeast states of Brazil. This study is the first to perform the molecular analysis of B. vogeli in Piauí. The results demonstrate that canine babesiosis is endemic in dogs sampled in Teresina and that PCR may be the method of choice to perform parasite screening in this region.

show abstract

“…Babesia rossi X 5 [16] Babesia canis X 5 [16] Ehrlichia canis X 5 [17] Ehrlichia chaffeensis X 5 [18] Anaplasma platys X 16 [19] Anaplasma phagocytophilum X 9 [20] Rickettsia conorii X 8 [21] Rickettsia africae X 8 [21] Rickettsia felis X X [22] Coxiella burnetti X 8 [23] Hepatozoon canis X 5…”

Section: Referencesmentioning

confidence: 99%

A community approach for pathogens and their arthropod vectors (ticks and fleas) in cats of sub-Saharan Africa

Madder¹,

Day

Schunack³

et al. 2022

Parasites Vectors

View full text Add to dashboard Cite

Background Arthropod-borne pathogens and their vectors are present throughout Africa. They have been well studied in livestock of sub-Saharan Africa, but poorly studied in companion animals. Given their socioeconomic importance, the African Small Companion Animal Network (AFSCAN), as part of the WSAVA Foundation, initiated a standardized multi-country surveillance study. Methods In six countries (Ghana, Kenya, Nigeria, Tanzania, Uganda, and Namibia) in both rural and urban settings, 160 infested cats were sampled to assess their ectoparasite community (ticks and fleas), as well as the micro-parasite prevalence within those ectoparasites (60 and 118 pools of ticks and fleas, respectively) and blood (276 cats, including 116 non-infested). Results Almost two thirds of all infested cats originated from Tanzania and Kenya. Despite the large macro-geographical variation, no consistent difference was found in ectoparasite diversity and numbers between East and West Africa. Far more flea-infested than tick-infested cats were found. The most dominant ectoparasite was Ctenocephalides felis. Among the ticks, the exophilic Haemaphysalis spp. were the commonest, including species that are not typically linked with companion animals (Haemaphysalis spinulosa and Haemaphysalis elliptica). The most prevalent pathogens found in the blood and fleas were Bartonella henselae and Mycoplasma haemofelis. In the ticks, the dog-associated Hepatozoon canis was most commonly found. A high degree of co-parasitism was found in all countries and habitats. Conclusions Our continent-wide standardized field study highlights the cat’s potential to serve as a reservoir of pathogens that can be transmitted to humans or livestock, especially when cats are expected to become more commonly kept in African villages and towns. Graphical Abstract

show abstract

Development and validation of a multiplex, real-time PCR assay for Babesia rossi and Babesia vogeli

Cited by 16 publications

References 57 publications

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

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

Molecular, serological, and parasitological detection of Babesia vogeli in dogs in the state of Piauí, Brazil

A community approach for pathogens and their arthropod vectors (ticks and fleas) in cats of sub-Saharan Africa

Contact Info

Product

Resources

About