Poedji Hastutiek and Mufasirin and Ferizka Fahmi Qurrota A’yuun and Willy Nara Putra (2018) Prospect of Antigenic Protein Whole Extract of Rhipicephalus sanguineus larvae for The Development of Anti-tick Vaccine in Dogs. Advances in Health Sciences Research (AHSR), 5. pp. 307-310. ISSN 978-94-6252-481-1

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Abstract

The aim of this research is to know antigenic protein of tick Rhipicephalus sanguineus and determine the humoral immune response in rabbits immunized with protein larvae of R. sanguineus which were expected to be used as a basic for the development of anti-tick vaccine. This research used larvae of ticks of Rhipicephalus sanguineus obtained from dog skin for making whole extract. Whole tick extract protein was obtained by sonication technique then the protein was analyzed using SDS-PAGE technique and continued with identification of antigenic proteins by Western blot technique. Antibodies obtained from the blood of rabbits immunized whole extract larvae of R. sanguineus. Blood sampling performed on day 14, 21, 28 and 35. Antibodies and antigens obtained are used for the Indirect ELISA test then read by an ELISA reader at a wavelength of 405 nm. The results of the ELISA reader in the form of OD values. OD values analysis by descriptive statistics using SPSS. Analysis result whole extract protein R. sanguineus with SDS-PAGE technique gained 3 protein bands with molecular weight of 95.3 kDa, 45.2 kDa and 24.4 kDa, respectively. The results of identification whole extract protein R. sanguineus by Western blot technique acquired 2 antigenic protein bands with molecular weight of 95.3 kDa and 45.2 kDa, respectively. The proteins are potential to be developed material of anti-tick vaccine. Statistical analysis showed OD values at blood sampling day 14, 21, 28 and 35 was 0.060 ± 0.009; 0.201 ± 0.038; 0.147 ± 0.025 and 0.296 ± 0.035. In the graph shown increasing OD values at days 21 and 35, while a decrease in OD values occurred on day 28. The conclusion of this study is immunization with whole extract of R. sanguineus larvae can cause humoral immune response in rabbits with the highest OD value on day 35. Keywords: Rhipicephalus sanguineus, Whole protein extract, SDS-PAGE, Western blot, Indirect ELISA

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