UNIVERSITAS AIRLANGGA



Detail Article

BIOSAINS

ISSN 1412-1433

Vol. 13 / No. 1 / Published : 2011-01

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Original Article :

Characterization and isolation of specific protein from excretory-secretory (es) material of l2 dormant of toxocara cati for the diagnostic development of toxocariasis by elisa tehnique

Author :

  1. Kusnoto*1
  2. Sri Subekti*2
  3. I Ketut Sudiana*3
  4. Soedarto*4
  1. Fakultas Kedokteran Hewan Universitas Airlangga
  2. Fakultas Kedokteran Hewan Universitas Airlangga
  3. Fakultas Kedokteran Universitas Airlangga
  4. Fakultas Kedokteran Universitas Airlangga

Abstract :

The objective of this study was to get the specific protein of T. cati that could be used as biomarker in kit diagnostic to toxocariasis diagnosis through antibody test by ELISA technique. This study was conducted by isolating and characterizis T. cati protein from secretory-excretory (ES) material of L2 dormant (L2D) by pure protein that was reactived with polyclonal antibody sera. For the first step, ES protein of L2D was identified by SDS-PAGE with coumsie blue staining. The second step, protein was transferred to nitrocellulose membrane by using polyclonal antibody of anti-L2D T. cati that was visualized through goat-anti rabbit conjugate and BCIP+NBT staining. The third step to decide the specific protein fractions based on the molecule weight (MW), and then the specific protein was isolated by elution technique. The fourth was to evaluate the antigenicity, sensitivity and specificity of pure protein by IgG-ELISA as a response against L2D T. cati immunization toward mice. The results of the study showed that: T. cati have more protein with varied molecular weight ranging from 8 to 200 kDa. The specific proteins of T. cati recognized by the anti-L2D T. cati sera are the proteins of MW 168, 140, 120, 80, 70, 42, 32, 30, 24, 18, 14, 12 dan 10 kDa. The antigenicity of protein of T. cati was higher than that of other worms (D_caninum and Ancylostoma spp.). The sensitivity of protein of T. cati in mice with immunization was 100%, the specificity was 87.5% and the false positive was 12.5%.

Keyword :

toxocariasis, , specificity, , diagnostic, , sensitivity, , ELISA,


References :

A bbas AK, Lichtman AH and Pober JS,(2000) Cellular and Mollecular Immunology. 4th ed. Philadelphia : Philadelphia Saunders Company

A bdel-Rahman EH and Abdel-Megeed KN,(2000) Molecular identity of major cross-reactive adult antigens in Fasciola gigantica, Toxocara vitulorum and Moniezia expanza. Egypt : Abstract. J. Egypt. Soc. Parasitol

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Dubinsky P, Akao N, Reiterova K and Konakova G,(2000) Comparison of the sensitive screening kit with two ELISA sets for detection of anti-Toxocara antibodies. Southeast Asian : Southeast Asian J Trop Med Public Health

Harlow E and Lane D,(1998) Production of monoklonal antibodies. In: Antibodies New York : A Laboratory Manual. Cold Spring Harbor Lab





Archive Article

Cover Media Content

Volume : 13 / No. : 1 / Pub. : 2011-01
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  2. Bioactivity Human Menopause Gonadotrophin (hmg) And Hmg Deglycosylated (hmgdg) Of Urine Of Postmenopause Woman Toward Cleavage Embryo Of Cattle In Vitro
  3. Extracellular Signal Regulated Kinase (erk1/2), Reseptor Estrogen (er) A, And Insuline Like Growth Factor I (igf-ir) Expressions On Osteoblast In Influencing Bone Density In The Female Young Rat (rattus Norvegicus) After Exercise Training
  4. The Role Of Endonuclease-g For Amniotic Cell Apoptosis Biomarker Determination In Pregnancies With Premature Rupture Of The Membrane
  5. The Microdeletion Of Rbm And Daz Genes In Male Partner Of Infertile Couple In Population With Inbreeding
  6. Compartion Study Between Animal Model Sopk And Normally Clycle Is Mmp-9, Timp-1 Enzym Activities And Collagen Expression (an Experiment Study In Rat)
  7. Characterization And Isolation Of Specific Protein From Excretory-secretory (es) Material Of L2 Dormant Of Toxocara Cati For The Diagnostic Development Of Toxocariasis By Elisa Tehnique

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