Development of a SYBR Green Real-Time Polymerase Chain Reaction Assay for Quantitative Detection of<i>Babesia Gibsoni</i>(Asian Genotype) DNA

Matsuu, Aya; Ono, Satoshi; Ikadai, Hiromi; Uchide, Tsuyoshi; Imamura, Saiki; Onuma, Misao; Okano, Shozo; Higuchi, Seiichi

doi:10.1177/104063870501700608

Cited by 35 publications

(24 citation statements)

References 21 publications

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“…This is in agreement with Matsuu et al (2005) who reported detection limit of light microscopy as approximately 0.001 % parasitaemia compared to PCR capable of detecting 9 parasites per ll of blood similar to 50 organisms/ml of blood reported by Birkenheuer et al (2003).…”

Section: Resultssupporting

confidence: 81%

Babesia infection in naturally exposed pet dogs from a north-eastern state (Assam) of India: detection by microscopy and polymerase chain reaction

Laha

Bhattacharjee²,

Sarmah³

et al. 2013

J Parasit Dis

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The objective of the study was to detect Babesia infections in pet dogs of a north-eastern state of India. The diagnostic efficacy of Babesia infection by polymerase chain reaction (PCR) technique has been compared with microscopy examination. For this, a total of 111 blood samples of pet dogs presented at clinical complex of the College of Veterinary Science, Guwahati, Assam with clinical signs suspected for Babesia infection subjected to the study. A total of 44 (39.63 %) dogs were diagnosed as positive for Babesia infections after microscopic examination. Among these, Babesia canis infection was diagnosed in 5 dogs (4.50 %) and B. gibsoni infection in 39 (35.13 %) dogs microscopically in Giemsa stained blood smears. Molecular diagnosis using PCR detected 63 (56.75 %) dogs positive for Babesia infection. Single infection with B. canis was found in 9 (8.10 %) dogs while B. gibsoni alone was detected in 3 (2.70 %) dogs. Mixed infections by both these species were detected in 51 (45.94 %) dogs. Overall, PCR detected 54 (48.64 %) dogs as B. gibsoni and 60 (54.05 %) dogs as B. canis positive.

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Section: Resultssupporting

confidence: 81%

Babesia infection in naturally exposed pet dogs from a north-eastern state (Assam) of India: detection by microscopy and polymerase chain reaction

Laha

Bhattacharjee²,

Sarmah³

et al. 2013

J Parasit Dis

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“…1). It is worthwhile mentioning that this region was encompassed in the primer target of two published B. gibsoni-specific polymerase chain reaction (PCR) methods, including both conventional PCR and real-time PCR [4,16]. In terms of the sequence divergence shown in Fig.…”

Section: Discussionmentioning

confidence: 99%

Sequence and Phylogenetic Analysis of the Thrombospondin-Related Adhesive Protein (TRAP) Gene of Babesia gibsoni Isolates from Dogs in Taiwan

et al. 2010

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ABSTRACT. The genetic diversity of Babesia gibsoni strains worldwide is currently poorly defined. The aim of the present study was to characterize B. gibsoni strains in naturally infected dogs in Taiwan using a combination of polymerase chain reaction (PCR) and sequence analysis of both 18S rDNA and the gene encoding thrombospondin-related adhesive protein (TRAP). Genomic DNA was extracted from 29 parasitemic dogs, and the target genes were separately amplified, sequenced and aligned with corresponding sequences available in GenBank. All 18S rDNA sequences (1,262 bp) amplified from the Taiwanese isolates were identical to each other and had very high similarity (99.9-100%) with previously reported B. gibsoni sequences. These results provide the first molecular evidence showing infection of dogs with B. gibsoni from Taiwan. On the other hand, a phylogenetic analysis based on the deduced amino acid sequence of the TRAP gene demonstrated that the Taiwanese isolates were closely related to strains previously identified from Okinawa Island, Japan, but genetically distinct from strains found on Honshu in Japan and Jeju Island in South Korea. The divergence of TRAP among the geographically dispersed strains examined in this study and others supports the conclusion that this gene is useful for molecular genotyping of B. gibsoni strains.

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“…The copy number of the B. gibsoni p18 gene in the peripheral blood was estimated using the real-time PCR method by Matsuu et al [18]. DNA was extracted from 200 µl of EDTA-anticoagulated whole blood samples using a commercial kit (QIAgen DNA Blood Mini Kit; Qiagen, Tokyo, Japan).…”

Section: Quantification Of B Gibsoni P18 Gene In the Blood By Real-tmentioning

confidence: 99%

Evaluation of Efficacy of Bruceine A, a Natural Quassinoid Compound Extracted from a Medicinal Plant, Bruced javanica, for Canine Babesiosis

Mizukami

Kawamura

subeki

et al. 2009

The Journal of Veterinary Medical Science

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ABSTRACT. Bruceine A, a natural quassinoid compound extracted from the dried fruits of Brucea javanica (L.) Merr., was evaluated for its antibabesial activity in vitro and in vivo. Bruceine A inhibited the in vitro growth of Babesia gibsoni in canine erythrocytes at lower concentration compared with the standard antibabesial drug diminazene aceturate and killed the parasites within 24 hr at a concentration of 25 nM. Oral administration of bruceine A at a dosage of 6.4 mg/kg/day for 5 days resulted in no clinical findings in a dog with normal ranges of hematological and biochemical values in the blood. Three dogs were infected with B. gibsoni and two of them were treated with bruceine A at a dosage of 6.4 mg/kg/day for 6 days from day 5 post-infection. An untreated dog developed typical acute babesiosis symptoms including severe anemia, high fever, and complete loss of appetite and movement. However, the two bruceine A-treated dogs maintained their healthy conditions throughout the experimental period of 4 weeks although complete elimination of parasites from the peripheral blood was not achieved and decreases in the packed cell volume and the erythrocyte and platelet counts were observed. Since natural quassinoid compounds have been used as traditional medicines for the treatment of various ailments including cancer and malaria, the present results suggest that bruceine A or other related compounds are potential candidates for the treatment of canine babesiosis.

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Development of a SYBR Green Real-Time Polymerase Chain Reaction Assay for Quantitative Detection ofBabesia Gibsoni(Asian Genotype) DNA

Cited by 35 publications

References 21 publications

Babesia infection in naturally exposed pet dogs from a north-eastern state (Assam) of India: detection by microscopy and polymerase chain reaction

Babesia infection in naturally exposed pet dogs from a north-eastern state (Assam) of India: detection by microscopy and polymerase chain reaction

Sequence and Phylogenetic Analysis of the Thrombospondin-Related Adhesive Protein (TRAP) Gene of Babesia gibsoni Isolates from Dogs in Taiwan

Evaluation of Efficacy of Bruceine A, a Natural Quassinoid Compound Extracted from a Medicinal Plant, Bruced javanica, for Canine Babesiosis

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