Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Acta Parasitologica


IMPACT FACTOR 2017: 1.039
5-year IMPACT FACTOR: 1.121

CiteScore 2018: 1.00

SCImago Journal Rank (SJR) 2018: 0.500
Source Normalized Impact per Paper (SNIP) 2018: 0.664

More options …
Volume 59, Issue 1

Issues

A PCR method targeting internal transcribed spacers: the simultaneous detection of Babesia bigemina and Babesia bovis in cattle

Junlong Liu
  • State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Xujiaping 1, Lanzhou, Gansu, 730046, P. R. China
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Guiquan Guan
  • State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Xujiaping 1, Lanzhou, Gansu, 730046, P. R. China
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Aihong Liu
  • State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Xujiaping 1, Lanzhou, Gansu, 730046, P. R. China
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Youquan Li
  • State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Xujiaping 1, Lanzhou, Gansu, 730046, P. R. China
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Hong Yin
  • State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Xujiaping 1, Lanzhou, Gansu, 730046, P. R. China
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Jianxun Luo
  • State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Xujiaping 1, Lanzhou, Gansu, 730046, P. R. China
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2014-02-26 | DOI: https://doi.org/10.2478/s11686-014-0222-6

Abstract

In this study, two pairs of oligonucleotide primers were designed according to the nucleotide sequence of the internal transcribed spacers (ITSs) of Babesia bigemina and B. bovis isolates from China. The primers were used in a multiplex PCR to detect parasite DNA in blood samples from cattle. There was no cross reactions with B. ovata, B. major, B. sp. Kashi, Theileria annulata, T. sergenti, T. sinensis or normal bovine DNA. The sensitivity of multiplex PCR assay was 1 pg and 10 pg DNA for B. bigemina and B. bovis, respectively. A total of 260 field blood samples collected from cattle in five provinces of China were analyzed by multiplex PCR and light microscopy. PCR testing revealed that 7.3% (19/260) and 5.8% (15/260) of cattle were positive for B. bigemina and B. bovis and 1.2% (3/260) of cattle were co-infected with B. bigemina and B. bovis. Using light microscopy, 2.3% (6/260) and 1.5% (4/260) of cattle were infected by B. bigemina and B. bovis, respectively, and no co-infection was found. The results showed that the multiplex PCR developed in the present study could be an alternative diagnostic tool for the detection of B. bovis and B. bigemina infection in cattle.

Keywords: Babesia bigemina; Babesia bovis; Multiplex PCR; ITS; ribosomal DNA

  • [1] Aktas M., Bendele K.G., Altay K., Dumanli N., Tsuji M., Holman P.J. 2007. Sequence polymorphism in the ribosomal DNA internal transcribed spacers differs among Theileria species. Veterinary Parasitology, 147, 221–230. DOI: 10.1016/j.vetpar.2007.04.007. http://dx.doi.org/10.1016/j.vetpar.2007.04.007CrossrefWeb of ScienceGoogle Scholar

  • [2] Alhassan A., Pumidonming W., Okamura M., Hirata H., Battsetseq B., Fujisaki K., Yokoyama N., Igarashi I. 2005. Development of a single-round and multiplex PCR method for the simultaneous detection of Babesia caballi and Babesia equi in horse blood. Veterinary Parasitology, 129, 43–49. DOI: 10.1016/j.vetpar.2004.12.018. http://dx.doi.org/10.1016/j.vetpar.2004.12.018CrossrefGoogle Scholar

  • [3] Allred D.R. 2003. Babesiosis: persistence in the face of adversity. Trends in Parasitology, 19, 51–55. http://dx.doi.org/10.1016/S1471-4922(02)00065-XCrossrefGoogle Scholar

  • [4] Almeria S., Castellà J., Ferrer D., Ortuño A., Estrada-Peña A., Gutiérrez J.F. 2001. Bovine piroplasms in Minorca (Balearic Islands, Spain): a comparison of PCR-based and light microscopy detection. Veterinary Parasitology, 99, 249–259. http://dx.doi.org/10.1016/S0304-4017(01)00464-2CrossrefGoogle Scholar

  • [5] Baneth G., Kenny M.J., Tasker S., Anug Y., Shkap V., Levy A., Shaw S.E. 2004. Infection with a proposed new subspecies of Babesia canis, Babesia canis subsp. present in domestic cat. Journal of Clinical Microbiology, 42, 99–105. http://dx.doi.org/10.1128/JCM.42.1.99-105.2004CrossrefGoogle Scholar

  • [6] Barros S.L., Madruga C.R., Araujo F.R., Menk C.F., Almeida M.A., Melo E.P.S., Kessler R.H. 2005. Serological survey of Babesia bovis, Babesia bigemina and Anaplasma marginale antibodies in cattle from the semi-arid region of Bahia, Brazil, by enzyme linked immunosorbent assays. Memórias do Instituto Oswaldo Cruz, 100, 513–517. DOI:/S0074-02762005000600003. http://dx.doi.org/10.1590/S0074-02762005000600003CrossrefGoogle Scholar

  • [7] Bishop J.P., Adams L.G. 1973. Combination thin and thick blood films for the detection of Babesia parasitemia. American Journal of Veterinary Research, 34, 1213–1214. Google Scholar

  • [8] Conrad P.A., Kjemtrup A.M., Carreno R.A., Thomford J., Wainwright K., Eberhard M., Quick R., Telford S.R.3rd, Herwaldt B.L. 2006. Description of Babesia duncani n. sp. (Apicomplexa: Babesiidae) from humans and its differentiation from other piroplasms. International Journal for Parasitology, 36, 779–789. DOI: 10.1016/j.ijpara.2006.03.008. http://dx.doi.org/10.1016/j.ijpara.2006.03.008CrossrefGoogle Scholar

  • [9] Figueroa J.V., Chieves L.P., Johnson G.S., Buening G.M. 1993. Multiplex polymerase chain reactionbased assay for the detection of Babesia bigemina, Babesia bovis and Anaplasma marginale DNA in bovine blood. Veterinary Parasitology, 50, 69–81. http://dx.doi.org/10.1016/0304-4017(93)90008-BCrossrefGoogle Scholar

  • [10] Fu Y.B., Lai Z.S. 1997. A report of buffalo babesiosis. Guangxi Journal of Animals Husbandry Veterinary Medicine, 3, 46–48 (In Chinese). Google Scholar

  • [11] García-Sanmartín J., Nagore D., García-Pérez A.L., Juste R.A., Hurtado A. 2006. Molecular diagnosis of Theileria and Babesia species infecting cattle in Northern Spain using reverse line blot macroarrays. BMC Veterinary Research, 2, 16. DOI: 10.1186/1746-6148-2-16. http://dx.doi.org/10.1186/1746-6148-2-16CrossrefGoogle Scholar

  • [12] Gubbels J.M., de Vos A.P., van der Weide M., Viseras J., Schouls L.M., de Vries E., Jongejan F. 1999. Simultaneous detection of bovine Theileria and Babesia species by reverse line blot hybridization. Journal of Clinical Microbiology, 37, 1782–1789. Google Scholar

  • [13] Holman P.J., Bendele K.G., Schoelkopf L., Jones-Witthuhn R.L., Jones S.O. 2003. Ribosomal RNA analysis of Babesia odocoilei isolates from farmed reindeer (Rangifer tarandus tarandus) and elk (Cervus elaphus canadensis) in Wisconsin. Parasitology Research, 91, 378–383. DOI: 10.1007/s00436-003-0984-5. http://dx.doi.org/10.1007/s00436-003-0984-5CrossrefGoogle Scholar

  • [14] Kho W.G., Chung J.Y., Sim E.J., Kim M.Y., Kim D.W., Jongwutiwes S., Tanabe K. 2003. A multiplex polymerase chain reaction for the differential diagnosis of Plasmodium falciparum and Plasmodium vivax. Parasitology International, 52, 229–236. http://dx.doi.org/10.1016/S1383-5769(03)00028-XCrossrefGoogle Scholar

  • [15] Kim C.M., Blanco L.B.C., Alhassan A., Iseki H., Yokoyama N., Xuan X., Igarashi I. 2008. Development of a Rapid Immunochromatographic Test for Simultaneous Serodiagnosis of Bovine Babesioses caused by Babesia bovis and Babesia bigemina. American Journal of Tropical Medicine and Hygiene, 78, 117–121. Google Scholar

  • [16] Liu A., Guan G., Du P., Gou H., Liu Z., Liu J., Ma M., Yang J., Li Y., Niu Q., Ren Q., Bai Q., Yin H., Luo J. 2012. Loop-mediated isothermal amplification (LAMP) method based on two species-specific primer sets for the rapid identification of Chinese Babesia bovis and B. bigemina. Parasitology International, 61, 658–663. DOI: 10.1016/j.parint.2012.07.004. http://dx.doi.org/10.1016/j.parint.2012.07.004CrossrefWeb of ScienceGoogle Scholar

  • [17] Liu J.L., Yin H., Liu G.Y., Guan G.Q., Ma M.L., Liu A.H., Liu Z.J., Li Y.Q., Ren Q.Y., Dang Z.S., Gao J.L., Zhao H.P., Luo J.X. 2008. Discrimination of Babesia major and Babesia ovata based on ITS1-5.8S-ITS2 region sequences of rRNA gene. Parasitology Research, 102, 709–713. DOI: 10.1007/s00436-007-0818-y. http://dx.doi.org/10.1007/s00436-007-0818-yWeb of ScienceGoogle Scholar

  • [18] Markoulatos P., Siafakas N., Moncany M. 2002. Multiplex polymerase chain reaction: a practical approach. Journal of Clinical Laboratory Analysis, 16, 47–51. http://dx.doi.org/10.1002/jcla.2058CrossrefGoogle Scholar

  • [19] McCosker P.J. 1981. The global importance of babesiosis. In: Ristic M., Kreier J.P. (Eds.), Babesiosis. Academic Press, New York, NY, 24 pp. Google Scholar

  • [20] Mclaughlin G.L., Edlind T.D., Ihler G.M. 1986. Detection of Babesia bovis using DNA hybridization. Journal of Protozoology, 33, 125–128. http://dx.doi.org/10.1111/j.1550-7408.1986.tb05571.xCrossrefGoogle Scholar

  • [21] Notomi T., Okayama H., Masubuchi H., Yonekawa T., Watanabe K., Amino N., Hase T. 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Research, 28, E63. http://dx.doi.org/10.1093/nar/28.12.e63CrossrefGoogle Scholar

  • [22] Oliveira M.C.S., Oliveira-Sequeira T.C.G., Regitano L.C.A., Alencar M.M., Néo T.A., Silva A.M., Oliveira H.N. 2008. Detection of Babesia bigemina in cattle of different genetic groups and in Rhipicephalus (Boophilus) microplus tick. Veterinary Parasitology, 155, 281–286. DOI: 10.1016/j.vetpar.2008.04.022. http://dx.doi.org/10.1016/j.vetpar.2008.04.022Web of ScienceCrossrefGoogle Scholar

  • [23] Passos L.M., Bell-Sakyi L., Brown C.G. 1998. Immunochemical characterization of in vitro culture-derived antigens of Babesia bovis and Babesia bigemina. Veterinary Parasitology, 76, 239–249. http://dx.doi.org/10.1016/S0304-4017(98)00095-8CrossrefGoogle Scholar

  • [24] Qin W.X. 2005. Diagnosis and therapy of bovine babesiosis. Today Animal Husbandry and Veterinary Medicine, 8, 10 (in Chinese). Google Scholar

  • [25] Salih D.A., El Hussein A.M., Seitzer U., Ahmed J.S. 2007. Epidemiological studies on tick-borne diseases of cattle in Central Equatoria State, Southern Sudan. Parasitology Research, 101, 1035–1044. DOI: 10.1007/s00436-007-0583-y. http://dx.doi.org/10.1007/s00436-007-0583-yCrossrefWeb of ScienceGoogle Scholar

  • [26] Taylor M.A., Coop R.L., Wall R.L. 2007. Veterinary Parasitology. Blackwell Publishing Ltd, 9600 Garsington Road, Oxford OX4 2DQ, UK, 104–105. Google Scholar

  • [27] Tenter A.M., Friedhoff K.T. 1986. Serodiagnosis of experimental and natural Babesia equi and B. caballi infections. Veterinary Parasitology, 20, 49–61. http://dx.doi.org/10.1016/0304-4017(86)90092-0CrossrefGoogle Scholar

  • [28] Yang Z.C. 1984. A research concerning a four foci babesiosis outbreak. Veterinary Science in China, 4, 36–37 (in Chinese). Google Scholar

About the article

Published Online: 2014-02-26

Published in Print: 2014-03-01


Citation Information: Acta Parasitologica, Volume 59, Issue 1, Pages 132–138, ISSN (Online) 1896-1851, DOI: https://doi.org/10.2478/s11686-014-0222-6.

Export Citation

© 2014 W. Stefański Institute of Parasitology, PAS. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[1]
Yongchang Li, Jixu Li, Bayin Chahan, Qingyong Guo, Yang Zhang, Paul Franck Adjou Moumouni, Seung-Hun Lee, Mingming Liu, Eloiza May Galon, Huanping Guo, Yang Gao, Ruiqi Song, Min Li, Maria Agnes Tumwebaze, Byamukama Benedicto, and Xuenan Xuan
Parasitology International, 2019, Page 101925
[2]
Mirza Omar Abdallah, Qingli Niu, Jifei Yang, Muhammad Adeel Hassan, Peifa Yu, Guiquan Guan, Ze Chen, Guangyuan Liu, Jianxun Luo, and Hong Yin
Journal of Parasitology, 2017, Volume 103, Number 3, Page 221
[3]
Junlong Liu, Guiquan Guan, Youquan Li, Aihong Liu, Jianxun Luo, and Hong Yin
Frontiers in Microbiology, 2017, Volume 8
[4]
Véronique Lehrter, Damien Jouet, Emmanuel Liénard, Anouk Decors, and Cécile Patrelle
Infection, Genetics and Evolution, 2016, Volume 46, Page 94
[5]
Ying Li, Pengpeng Liu, Chengmin Wang, Gang Chen, Ming Kang, Daoxin Liu, Zengkui Li, Hongxuan He, Ying Dong, and Yanming Zhang
Journal of Wildlife Diseases, 2015, Volume 51, Number 4, Page 872

Comments (0)

Please log in or register to comment.
Log in