Rodolfo A Verdida

Publications (5)7.68 Total impact

• Article: Development of two immunochromatographic tests for the serodiagnosis of bovine babesiosis.

  Chulmin Kim, Andy Alhassan, Rodolfo A Verdida, Naoaki Yokoyama, Xuenan Xuan, Kozo Fujisaki, Shin-Ichiro Kawazu, Ikuo Igarashi
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  ABSTRACT: In this study, we developed two immunochromatographic tests (ICTs), which are nitrocellulose membrane-based immunoassays for the convenient and rapid serodiagnosis of bovine babesiosis caused by Babesia bovis (BoICT) and Babesia bigemina (BiICT). The efficacy of two ICTs was evaluated using 13 positive sera from experimentally infected cattle with B. bovis or B. bigemina. Clear results showed that the BoICT and ELISA detected antibodies in sera collected from 14 to 93 days post-infection, while BiICT and ELISA detected from 13 to 274 days post-infection. In additon, non-infected cattle, Neospora caninum, and Cryptosporidium parvum were negative in two ICTs. To evaluate the field utility of the ICTs, we tested 186 field bovine sera collected from cattle living in Yanbian (China) and Mato Grosso do Sul (Brazil). The results of ICTs were compared to those of classical serodiagnostic methods, enzyme-linked immunosorbent assay (ELISA) and the indirect immunofluorescence assay (IFAT). The overall concordances of BoICT were determined as 92.5 and 90.3% when the results of ELISA and IFAT were set as the reference standards, respectively. In contrast, those of BiICT showed 96.8 and 92.5% relative to the results of standard ELISA and IFAT, respectively. Conventional and rapid diagnosic devices for bovine babesiosis may provide a valuable tool in clinical and field applications.
  Veterinary Parasitology 10/2007; 148(2):137-43. · 2.58 Impact Factor
• Source

  Available from: obihiro.ac.jp

  Article: Clinical usefulness of antibodies against Babesia gibsoni detected by ELISA with recombinant P50.

  Takako Miyama, Hisashi Inokuma, Kazuhito Itamoto, Masaru Okuda, Rodolfo A Verdida, Xuenan Xuan
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  ABSTRACT: The clinical usefulness of antibodies against Babesia gibsoni detected by ELISA with recombinant P50 was examined in dogs in an area where B. gibsoni infection was endemic. Only 8 among 14 dogs with acute type B. gibsoni infection without a previous history of infection were positive. This high percentage of false-negative results is thought to be a weak point of ELISA as a diagnostic tool. However, 14 other anemic dogs with a confirmed history of B. gibsoni infection were all positive, thus confirming the higher sensitivity of ELISA in detecting a history of infection.
  Journal of Veterinary Medical Science 01/2007; 68(12):1371-3. · 0.85 Impact Factor
• Article: Immunochromatographic test for simultaneous serodiagnosis of Babesia caballi and B. equi infections in horses.

  Xiaohong Huang, Xuenan Xuan, Rodolfo A Verdida, Shoufa Zhang, Naoaki Yokoyama, Longshan Xu, Ikuo Igarashi
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  ABSTRACT: An immunochromatographic test for the simultaneous detection of Babesia caballi- and B. equi-specific antibodies (BceICT) was developed using a recombinant B. caballi 48-kDa rhoptry protein (rBc48) and a recombinant truncated B. equi merozoite antigen 2 (rEMA-2t). An evaluation of the ability of the BceICT to detect antibodies in sera from uninfected horses and experimentally infected horses showed high sensitivities and specificities of 83.3% (10/12 sera) and 92.9% (52/56 sera), respectively, for the anti-B. caballi antibody and 94.1% (16/17 sera) and 88.2% (45/51 sera), respectively, for the anti-B. equi antibody. Results from the detection of antibodies in field-collected sera indicated that the BceICT results corresponded with those of enzyme-linked immunosorbent assays (ELISA), showing 91.8% correspondence (67/73 sera) for B. caballi and 95.9% correspondence (70/73 sera) for B. equi, and that the BceICT results also corresponded with the ICT for B. caballi and for B. equi, both of which were 98.2% (55/56 sera). The comparable results of the ICT and ELISA and the simplicity and rapidity of the performance of the ICT suggest that the BceICT would be a feasible test for the simultaneous serodiagnosis of both agents of equine babesiosis in the field.
  Clinical and Vaccine Immunology 06/2006; 13(5):553-5. · 2.55 Impact Factor
• Source

  Available from: Malaika Watanabe

  Article: Epidemiological survey of Babesia gibsoni infection in dogs in eastern Japan.

  Takako Miyama, Yoshimi Sakata, Yojiro Shimada, Shoji Ogino, Malaika Watanabe, Kazuhito Itamoto, Masaru Okuda, Rodolfo A Verdida, Xuenan Xuan, Hideyuki Nagasawa, Hisashi Inokuma
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  ABSTRACT: To determine the distribution of Babesia gibsoni infection in dogs in the eastern part of Japan, an epidemiological survey of dogs suspected of having B. gibsoni infection was attempted using the polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Thirty-five of 115 such dogs (30.4%) were positive by PCR and/or ELISA. The 35 positive dogs consisted of 28 Tosa dogs, 4 American Pit Bull Terriers, and 3 mongrel dogs in Aomori, Fukushima, Ibaraki, Gunma, Chiba, Tokyo, Kanagawa, and Nagano Prefectures. The positive dogs had a significantly lower rate of tick exposure and a higher rate of bites by other dogs. Twenty-two of 35 B. gibsoni-positive dogs were infected with hemoplasma, and the rate of infection was significantly higher than that of B. gibsoni-negative dogs.
  Journal of Veterinary Medical Science 06/2005; 67(5):467-71. · 0.85 Impact Factor
• Article: Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50.

  Rodolfo A Verdida, Olgga A Hara, Xuenan Xuan, Shinya Fukumoto, Ikuo Igarashi, Shoufa Zhang, Junyan Dong, Hisashi Inokuma, Hidenori Kabeya, Yukita Sato, Tadaaki Moritomo, Soichi Maruyama, Florencia Claveria, Hideyuki Nagasawa
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  ABSTRACT: The surface antigen P50 of Babesia gibsoni is an important candidate for the development of a diagnostic reagent for canine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding truncated P50 (P50t) lacking a signal peptide and C-terminal hydrophobic regions were cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). More than 90% portion of the GST-P50t was expressed as a soluble form, in contrast with GST-P50f (full-length), which was completely expressed as an insoluble form. This result indicates that removal of the hydrophobic signal peptide and C-terminus had dramatically improved its hydrophilicity. The purified GST-P50t was tested in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to B. gibsoni in dogs. The ELISA with GST-P50t clearly differentiated between B. gibsoni-infected dog sera and uninfected dog sera. In addition, the ELISA detected no cross-reactivity with sera from dogs experimentally infected with the closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi. Field serum samples collected from dogs in Japan and China were examined for the diagnosis of B. gibsoni infection by using the ELISA. 14.5% (9/62), 5.8% (7/120), and 5.4% (2/37) of tested samples were positive for dogs from Okinawa, Yamaguchi, and Osaka prefectures, Japan, respectively. On the other hand, 4.8% (2/41) of tested samples were positive for dogs from Nanjing, China. These results suggest that the GST-P50t could be a reliable reagent for practical use in ELISA for the serodiagnosis of canine piroplasmosis caused by B. gibsoni.
  Journal of Veterinary Medical Science 01/2005; 66(12):1517-21. · 0.85 Impact Factor