- INTRODUCTION
- MATERIALS AND METHODS
- Serum.
- Immunoscreening of cDNA expression library.
- Southern blotting.
- Expression and purification of recombinant BgSA1.
- Preparation of rabbit and mouse sera against BgSA1.
- Indirect fluorescent antibody test and confocal laser microscopic observation.
- Western blotting.
- ELISA.
- Double-antibody sandwich ELISA.
- Nucleotide sequence accession number.
- RESULTS
- Identification of the BgSA1 gene.
- Characterization of the BgSA1 gene.
- Expression of BgSA1 in E. coli.
- Identification of the native BgSA1 of B. gibsoni.
- Evaluation of recombinant BgSA1 in an ELISA for the detection of a specific antibody.
- Evaluation of double-antibody sandwich ELISA for the detection of circulating BgSA1.
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Boozer AL, Macintire DK, 2003. Canine babesiosis. Vet Clin North Am Small Anim Pract 33 :855–904.
-
2
Casapulla R, Baldi L, Avallone V, Sannino R, Pazzanese L, Mizzoni V, 1998. Canine piroplasmosis due to Babesia gibsoni: Clinical and morphological aspects. Vet Rec 142 :168–169.
-
3
Wozniak EJ, Barr BC, Thomford JW, Yamane I, McDonough SP, Moore PF, Naydan D, Robinson TW, Conrad PA, 1997. Clinical, anatomic, and immunopathologic characterization of Babesia gibsoni infection in the domestic dog (Canis familiaris). J Parasitol 83 :692–699.
-
4
Macintire DK, Boudreaux MK, West GD, Bourne C, Wright JC, Conrad PA, 2002. Babesia gibsoni infection among dogs in the southeastern United States. J Am Vet Med Assoc 220 :325–329.
-
5
Muhlnickel CJ, Jefferies R, Ryan UM, Irwin PJ, 2002. Babesia gibsoni infection in three dogs in Victoria. Aust Vet J 8 :606–610.
-
6
Zhou J, Mulenga A, Yamasaki M, Ohashi K, Maede Y, Onuma M, 2002. Babesia gibsoni: Molecular cloning and characterization of Rab6 and Rab11 homologues. Exp Parasitol 101 :210–214.
-
7
Adachi K, Ueno C, Makimura S, 1993. Immunosuppression in dogs naturally infected with Babesia gibsoni.J Vet Med Sci 55 :503–505.
-
8
Inokuma H, Yoshizaki Y, Matsumoto K, Okuda M, Onishi T, Nakagome K, Kosugi R, Hirakawa M, 2004. Molecular survey of Babesia infection in dogs in Okinawa, Japan. Vet Parasitol 121 :341–346.
-
9
Miyama T, Sakata Y, Shimada Y, Ogino S, Watanabe M, Itamoto K, Okuda M, Verdida A, Xuan X, Nagasawa H, Inokuma H, 2005. Epidemiological survey of Babesia gibsoni infection in dogs in eastern Japan. J Vet Med Sci 67 :467–471.
-
10
Fukumoto S, Xuan X, Shigeno S, Kimbita E, Igarashi I, Nagasawa H, Fujisaki K, Mikami T, 2001. Development of a polymerase chain reaction method for diagnosing Babesia gibsoni infection in dogs. J Vet Med Sci 63 :977–981.
-
11
Fukumoto S, Sekine Y, Xuan X, Igarashi I, Sugimoto C, Nagasawa H, Fujisaki K, Mikami T, Suzuki N, 2004. Serodiagnosis of canine Babesia gibsoni infection by enzyme-linked immunosorbent assay with recombinant P50 expressed in Escherichia coli.J Parasitol 90 :387–391.
-
12
Verdida RA, Hara OA, Xuan X, Fukumoto S, Igarashi I, Zhang S, Dong J, Inokuma H, Kabeya H, Sato Y, Moritomo T, Maruyama S, Claveria F, Nagasawa H, 2004. Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50. J Vet Med Sci 66 :1517–1521.
-
13
Yamane I, Thomford JW, Gardner IA, Dubey JP, Levy M, Conrad PA, 1993. Evaluation of the indirect fluorescent antibody test for diagnosis of Babesia gibsoni infections in dogs. Am J Vet Res 54 :1579–1584.
-
14
Fukumoto S, Xuan X, Igarashi I, Zhang S, Mugisha J, Ogata T, Nagasawa H, Fujisaki K, Suzuki N, Mikami T, 2000. Morphological changes of Babesia gibsoni proliferated in canine red blood cell-substituted severe combined immunodeficiency mice. J Parasitol 86 :956–959.
-
15
Fukumoto S, Xuan X, Nishikawa Y, Inoue N, Igarashi I, Nagasawa H, Fujisaki K, Mikami T, 2001b. Identification and expression of a 50-kilodalton surface antigen of Babesia gibsoni and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay. J Clin Microbiol 39 :2603–2609.
-
16
Bork S, Okamura M, Boonchit S, Hirata H, Yokoyama N, Igarashi I, 2004. Identification of Babesia bovis L-lactate dehydrogenase as a potential chemotherapeutical target against bovine babesiosis. Mol Biochem Parasitol 2 :165–172.
-
17
Xuan X, Maeda K, Mikami T, Otsuka H, 1996. Characterization of canine herpesvirus glycoprotein C expressed in insect cells. Virus Res 46 :57–64.
-
18
Xuan X, Larsen A, Ikadai K, Tanaka T, Igarashi I, Nagasawa H, 2001. Expression of Babesia equi merozoite antigen 1 in insect cells by recombinant baculovirus and evaluation of its diagnostic potential in an enzyme-linked Immunosorbent assay. J Clin Microbiol 39 :705–709.
-
19
Ikadai H, Tanaka H, Shibahara N, Matsuu A, Uechi M, Itoh N, Oshiro S, Kudo N, Igarashi I, Oyamada T, 2004. Molecular evidence of infections with Babesia gibsoni parasites in Japan and evaluation of the diagnostic potential of a loop-mediated isothermal amplification method. J Clin Microbiol 42 :2465–2469.
-
20
Homer MJ, Lodes MJ, Reynolds LD, Zhang Y, Douglass JF, McNeill PD, Houghton RL, Persing DH, 2003. Identification and characterization of putative secreted antigens from Babesia microti.J Clin Microbiol 41 :723–729.
-
21
Alifrangis M, Ihristensen FT, Jorgensen CS, Ronn AM, Weng JE, Chen M, Bygjerg IC, Sirawaraporn W, Palarasah Y, Koch C, 2004. Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum.Malar J 12 :3–16.
-
22
De Arruda ME, Collins KM, Hochberg LP, Ryan PR, Wirtz RA, Ryan JR, 2004. Quantitative determination of sporozoites and circumsporozoite antigen in mosquitoes infected with Plasmodium falciparum or P. vivax.Ann Trop Med Parasitol 98 :121– 127.
-
23
Noedl H, Wernsdorfer WH, Miller RS, Wongsrichanalai C, 2002. Histidine-rich protein II: a novel approach to antimalarial drug susceptibility testing. Antimicrob Agents Chemother 46 :1658–1664.
-
24
Dondorp AM, Desakorn V, Pongtavornpinyo W, Sahassananda D, Silamut K, Chotivanich K, Newton PN, Pitisuttithum P, Smithyman AM, White NJ, Day NP, 2005. Estimation of the total parasite biomass in acute falciparum malaria from plasma PfHRP2. PLoS Med 8 :e204.
-
25
Benito A, Carmena D, 2005. Double-antibody sandwich ELISA using biotinylated antibodies for the detection of Echinococcus granulosus coproantigens in dogs. Acta Trop 95 :9–15.
-
26
Khusmith S, Intapan P, Tharavanij S, Tuntrakul S, Indravijit KA, Bunnag D, 1992. Two-site sandwich ELISA for detection of Plasmodium vivax blood stage antigens using monoclonal and polyclonal antibodies. Southeast Asian J Trop Med Public Health 4 :745–751.
-
27
Beniwal RP, Nichani AK, Rakha NK, Sharma RD, Sarup S, 1997. An immunization trial with in vitro produced Babesia bigemina exoantigens. Trop Anim Health Prod 29 :124s–126s.
-
28
Lewis BD, Penzhorn BL, Lopez Rebollar LM, 1995. Immune responses to South African Babesia canis and the development of a preliminary vaccine. J S Afr Vet Assoc 66 :61–65.
-
29
Schettes TP, Kleuskens JA, Scholte NC, Pasman JW, Bos HJ, 1994. Vaccination of dogs against Babesia canis infection using antigens from culture supernatants with emphasis on clinical babesiosis. Vet Parasitol 52 :219–233.
-
30
Valentin A, Precigout E, L’Hostis M, Carcy B, Gorenflot A, Schrevel J, 1993. Cellular and humoral immune responses induced in cattle by vaccination with Babesia divergens culture-derived exoantigens correlate with protection. Infect Immun 61 :734–741.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 740 | 670 | 14 |
| Full Text Views | 248 | 14 | 2 |
| PDF Downloads | 45 | 8 | 0 |
IDENTIFICATION OF A NOVEL GENE ENCODING A SECRETED ANTIGEN 1 OF BABESIA GIBSONI AND EVALUATION OF ITS USE IN SERODIAGNOSIS
HONGLIN JIA
HONGLIN JIA
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan; School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan
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JINLIN ZHOU
JINLIN ZHOU
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan; School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan
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HIROMI IKADAI
HIROMI IKADAI
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan; School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan
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AYA MATSUU
AYA MATSUU
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan; School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan
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HIROSHI SUZUKI
HIROSHI SUZUKI
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan; School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan
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IKUO IGARASHI
IKUO IGARASHI
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan; School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan
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KOZO FUJISAKI
KOZO FUJISAKI
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan; School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan
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XUENAN XUAN
XUENAN XUAN
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan; School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan
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Serum from a dog immunized with blood plasma from a B. gibsoni-infected dog, putatively containing secreted antigens, was used to screen a cDNA expression library. A novel gene encoding BgSA1 was identified from the isolated clones. The serum raised in mice immunized with the recombinant BgSA1 expressed in Escherichia coli could recognize a native parasite protein with a molecular mass of 59 kDa. Comparing with the previously established ELISA with recombinant P50 as antigen, the ELISA with recombinant BgSA1 as the antigen was more sensitive when they were used to detect field samples. Moreover, a sandwich ELISA with anti-BgSA1 antibodies could detect the circulating BgSA1 in a serial blood plasma from a dog experimentally infected with B. gibsoni. These results indicated that BgSA1 could be a useful target for the development of a diagnostic test for the detection of specific antibodies and circulating antigens.
Author Notes
- INTRODUCTION
- MATERIALS AND METHODS
- Serum.
- Immunoscreening of cDNA expression library.
- Southern blotting.
- Expression and purification of recombinant BgSA1.
- Preparation of rabbit and mouse sera against BgSA1.
- Indirect fluorescent antibody test and confocal laser microscopic observation.
- Western blotting.
- ELISA.
- Double-antibody sandwich ELISA.
- Nucleotide sequence accession number.
- RESULTS
- Identification of the BgSA1 gene.
- Characterization of the BgSA1 gene.
- Expression of BgSA1 in E. coli.
- Identification of the native BgSA1 of B. gibsoni.
- Evaluation of recombinant BgSA1 in an ELISA for the detection of a specific antibody.
- Evaluation of double-antibody sandwich ELISA for the detection of circulating BgSA1.
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Boozer AL, Macintire DK, 2003. Canine babesiosis. Vet Clin North Am Small Anim Pract 33 :855–904.
-
2
Casapulla R, Baldi L, Avallone V, Sannino R, Pazzanese L, Mizzoni V, 1998. Canine piroplasmosis due to Babesia gibsoni: Clinical and morphological aspects. Vet Rec 142 :168–169.
-
3
Wozniak EJ, Barr BC, Thomford JW, Yamane I, McDonough SP, Moore PF, Naydan D, Robinson TW, Conrad PA, 1997. Clinical, anatomic, and immunopathologic characterization of Babesia gibsoni infection in the domestic dog (Canis familiaris). J Parasitol 83 :692–699.
-
4
Macintire DK, Boudreaux MK, West GD, Bourne C, Wright JC, Conrad PA, 2002. Babesia gibsoni infection among dogs in the southeastern United States. J Am Vet Med Assoc 220 :325–329.
-
5
Muhlnickel CJ, Jefferies R, Ryan UM, Irwin PJ, 2002. Babesia gibsoni infection in three dogs in Victoria. Aust Vet J 8 :606–610.
-
6
Zhou J, Mulenga A, Yamasaki M, Ohashi K, Maede Y, Onuma M, 2002. Babesia gibsoni: Molecular cloning and characterization of Rab6 and Rab11 homologues. Exp Parasitol 101 :210–214.
-
7
Adachi K, Ueno C, Makimura S, 1993. Immunosuppression in dogs naturally infected with Babesia gibsoni.J Vet Med Sci 55 :503–505.
-
8
Inokuma H, Yoshizaki Y, Matsumoto K, Okuda M, Onishi T, Nakagome K, Kosugi R, Hirakawa M, 2004. Molecular survey of Babesia infection in dogs in Okinawa, Japan. Vet Parasitol 121 :341–346.
-
9
Miyama T, Sakata Y, Shimada Y, Ogino S, Watanabe M, Itamoto K, Okuda M, Verdida A, Xuan X, Nagasawa H, Inokuma H, 2005. Epidemiological survey of Babesia gibsoni infection in dogs in eastern Japan. J Vet Med Sci 67 :467–471.
-
10
Fukumoto S, Xuan X, Shigeno S, Kimbita E, Igarashi I, Nagasawa H, Fujisaki K, Mikami T, 2001. Development of a polymerase chain reaction method for diagnosing Babesia gibsoni infection in dogs. J Vet Med Sci 63 :977–981.
-
11
Fukumoto S, Sekine Y, Xuan X, Igarashi I, Sugimoto C, Nagasawa H, Fujisaki K, Mikami T, Suzuki N, 2004. Serodiagnosis of canine Babesia gibsoni infection by enzyme-linked immunosorbent assay with recombinant P50 expressed in Escherichia coli.J Parasitol 90 :387–391.
-
12
Verdida RA, Hara OA, Xuan X, Fukumoto S, Igarashi I, Zhang S, Dong J, Inokuma H, Kabeya H, Sato Y, Moritomo T, Maruyama S, Claveria F, Nagasawa H, 2004. Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50. J Vet Med Sci 66 :1517–1521.
-
13
Yamane I, Thomford JW, Gardner IA, Dubey JP, Levy M, Conrad PA, 1993. Evaluation of the indirect fluorescent antibody test for diagnosis of Babesia gibsoni infections in dogs. Am J Vet Res 54 :1579–1584.
-
14
Fukumoto S, Xuan X, Igarashi I, Zhang S, Mugisha J, Ogata T, Nagasawa H, Fujisaki K, Suzuki N, Mikami T, 2000. Morphological changes of Babesia gibsoni proliferated in canine red blood cell-substituted severe combined immunodeficiency mice. J Parasitol 86 :956–959.
-
15
Fukumoto S, Xuan X, Nishikawa Y, Inoue N, Igarashi I, Nagasawa H, Fujisaki K, Mikami T, 2001b. Identification and expression of a 50-kilodalton surface antigen of Babesia gibsoni and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay. J Clin Microbiol 39 :2603–2609.
-
16
Bork S, Okamura M, Boonchit S, Hirata H, Yokoyama N, Igarashi I, 2004. Identification of Babesia bovis L-lactate dehydrogenase as a potential chemotherapeutical target against bovine babesiosis. Mol Biochem Parasitol 2 :165–172.
-
17
Xuan X, Maeda K, Mikami T, Otsuka H, 1996. Characterization of canine herpesvirus glycoprotein C expressed in insect cells. Virus Res 46 :57–64.
-
18
Xuan X, Larsen A, Ikadai K, Tanaka T, Igarashi I, Nagasawa H, 2001. Expression of Babesia equi merozoite antigen 1 in insect cells by recombinant baculovirus and evaluation of its diagnostic potential in an enzyme-linked Immunosorbent assay. J Clin Microbiol 39 :705–709.
-
19
Ikadai H, Tanaka H, Shibahara N, Matsuu A, Uechi M, Itoh N, Oshiro S, Kudo N, Igarashi I, Oyamada T, 2004. Molecular evidence of infections with Babesia gibsoni parasites in Japan and evaluation of the diagnostic potential of a loop-mediated isothermal amplification method. J Clin Microbiol 42 :2465–2469.
-
20
Homer MJ, Lodes MJ, Reynolds LD, Zhang Y, Douglass JF, McNeill PD, Houghton RL, Persing DH, 2003. Identification and characterization of putative secreted antigens from Babesia microti.J Clin Microbiol 41 :723–729.
-
21
Alifrangis M, Ihristensen FT, Jorgensen CS, Ronn AM, Weng JE, Chen M, Bygjerg IC, Sirawaraporn W, Palarasah Y, Koch C, 2004. Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum.Malar J 12 :3–16.
-
22
De Arruda ME, Collins KM, Hochberg LP, Ryan PR, Wirtz RA, Ryan JR, 2004. Quantitative determination of sporozoites and circumsporozoite antigen in mosquitoes infected with Plasmodium falciparum or P. vivax.Ann Trop Med Parasitol 98 :121– 127.
-
23
Noedl H, Wernsdorfer WH, Miller RS, Wongsrichanalai C, 2002. Histidine-rich protein II: a novel approach to antimalarial drug susceptibility testing. Antimicrob Agents Chemother 46 :1658–1664.
-
24
Dondorp AM, Desakorn V, Pongtavornpinyo W, Sahassananda D, Silamut K, Chotivanich K, Newton PN, Pitisuttithum P, Smithyman AM, White NJ, Day NP, 2005. Estimation of the total parasite biomass in acute falciparum malaria from plasma PfHRP2. PLoS Med 8 :e204.
-
25
Benito A, Carmena D, 2005. Double-antibody sandwich ELISA using biotinylated antibodies for the detection of Echinococcus granulosus coproantigens in dogs. Acta Trop 95 :9–15.
-
26
Khusmith S, Intapan P, Tharavanij S, Tuntrakul S, Indravijit KA, Bunnag D, 1992. Two-site sandwich ELISA for detection of Plasmodium vivax blood stage antigens using monoclonal and polyclonal antibodies. Southeast Asian J Trop Med Public Health 4 :745–751.
-
27
Beniwal RP, Nichani AK, Rakha NK, Sharma RD, Sarup S, 1997. An immunization trial with in vitro produced Babesia bigemina exoantigens. Trop Anim Health Prod 29 :124s–126s.
-
28
Lewis BD, Penzhorn BL, Lopez Rebollar LM, 1995. Immune responses to South African Babesia canis and the development of a preliminary vaccine. J S Afr Vet Assoc 66 :61–65.
-
29
Schettes TP, Kleuskens JA, Scholte NC, Pasman JW, Bos HJ, 1994. Vaccination of dogs against Babesia canis infection using antigens from culture supernatants with emphasis on clinical babesiosis. Vet Parasitol 52 :219–233.
-
30
Valentin A, Precigout E, L’Hostis M, Carcy B, Gorenflot A, Schrevel J, 1993. Cellular and humoral immune responses induced in cattle by vaccination with Babesia divergens culture-derived exoantigens correlate with protection. Infect Immun 61 :734–741.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 740 | 670 | 14 |
| Full Text Views | 248 | 14 | 2 |
| PDF Downloads | 45 | 8 | 0 |