Pengembangan Biosensor Penyakit Surra (Trypanosoma evansi) berbasis Protein dengan Metode Differential Pulse Voltammetry

A H Wardhana, H Munawar, D H Sawitri, R Maryam

Abstract

Trypanosoma evansi masih menjadi penyebab utama penyakit trypanosomiasis di Indonesia, yang dikenal dengan nama Surra. Namun demikian, kelemahan dalam mendiagnosa yang tepat dari Card Agglutination Test for Trypanosomiasis/T. evansi (CATT/T. evansi), ELISA and PCR menyebabkan pengendalian surra di lapang masih menghadapi kendala. Untuk itu, diperlukan alternatif piranti diagnosis yang cepat, akurat dan mudah diaplikasikan, yaitu biosensor. Penelitian ini bertujuan untuk mengetahui kinerja biosensor surra (Trypanosoma evansi) dalam mengidentifikasi sampel serum positif dan negatif. Sumber protein diisolasi dari T. evansi yang dipropagasi. Protein diimobilisasi dengan reaksi carbodiimide pada permukaan sensor (elektrode) karbon sebagai material yang memiliki respon terrendah terhadap serum/antibodi anti T. evansi (persentase arus listrik/elektron < 90). Metode yang digunakan pada pengembangan biosensor ini adalah differential pulse voltammetry (DPV). Data yang diperoleh dinormalisasi dan dianalisis menggunakan program PSTrace 5.8. Hasil penelitian membuktikan bahwa positif dan negatif palsu sensor dideteksi pada pengenceran serum 2 dan 64 kali. Serum positif yang dianalisis dengan metode DPV (kisaran potential listrik – 0.2 - +1.0 V dengan tingkat kecepatan pembacaan 0,1 Vs-1) menunjukkan bahwa slope serum positif (1,84) lebih tinggi dibandingkan dengan sampel negatif (1,01) yang mengindikasikan sensitivitas biosensor terhadap Surra (T. evansi). Hasil ini dikonfirmasi dengan melakukan pengujian serologis pada sampel menggunakan (Card Agglutination Test for Trypanosomiasis/T. evansi – CATT/T. evansi). Hasil penelitian ini menunjukkan bahwa biosensor surra/T. evansi berbasis protein merupakan piranti yang menjanjikan untuk dikembangkan dalam mendeteksi surra pada ternak.

 Keywords

Protein; DPV; biosensor; Trypanosoma evansi; surra

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References

Campuzano S, Yáñez-Sedeño P, Pingarrón JM. 2017. Electrochemical Biosensing for the Diagnosis of Viral Infections and Tropical Diseases. ChemElectroChem. 4:753–777.

Castilho M, Laube T, Yamanaka H, Alegret S, Pividori MI. 2011. Magneto immunoassays for plasmodium falciparum histidine-rich protein 2 related to malaria based on magnetic nanoparticles. Anal Chem. 83:5570–5577.

Dewi R, Wardhana A, Soejoedono R, Mulatsih S. 2019. Evaluation of surra treatment srategies attacking horse and buffaloes in East Sumba District, Nusa Tenggara Timur Province of Indonesia (2010-2016). Jurnal Ilm:39–48.

Dewi R, Wardhana A, Soejoedono R, Mulatsih S, Basri C. 2019. Economic analysis for selection of diagnostic methods against surra in Buffalo on East Sumba Island, Indonesia. Adv Heal Sci Res. 19:115–119.

Dridi F, Marrakchi M, Gargouri M, Garcia-Cruz A, Dzyadevych S, Vocanson F, Saulnier J, Jaffrezic-Renault N, Lagarde F. 2015. Thermolysin entrapped in a gold nanoparticles/polymer composite for direct and sensitive conductometric biosensing of ochratoxin A in olive oil. Sensors Actuators, B Chem [Internet]. 221:480–490. Available from: https://dx.doi.org/10.1016/j.snb.2015.06.120

Ferreira AAP, Colli W, Da Costa PI, Yamanaka H. 2005. Immunosensor for the diagnosis of Chagas’ disease. Biosens Bioelectron. 21:175–181.

Galal A, Atta NF, El-Ads EH. 2012. Probing cysteine self-assembled monolayers over gold nanoparticles - Towards selective electrochemical sensors. Talanta [Internet]. 93:264–273. Available from: https://dx.doi.org/10.1016/j.talanta.2012.02.032

Garcia MFK do. S, Andrade CAS, de Melo CP, Gomes DS, Silva LG, Dias R V., Balbino VQ, Oliveira MDL. 2016. Impedimetric sensor for Leishmania infantum genome based on gold nanoparticles dispersed in polyaniline matrix. J Chem Technol Biotechnol. 91:2810–2816.

Laha R, Sasmal N, Bandyopadhyay S. 2008. Comparative polypeptide profiles of whole cell lysate antigens of Trypanosoma evansi isolated from three different hosts of eastern India. J Protozool Res. 18:11–16.

Luckins A. 1992. Methods for diagnosis of Trypanosomiasis in livestock. [place unknown].

Martins BR, Barbosa YO, Andrade CMR, Pereira LQ, Simão GF, de Oliveira CJ, Correia D, Oliveira RTS, da Silva M V., Silva ACA, et al. 2020. Development of an Electrochemical Immunosensor for Specific Detection of Visceral Leishmaniasis Using Gold-Modified Screen-Printed Carbon Electrodes. Biosensors. 10:81.

Merli D, Ferrari C, Cabrini E, Dacarro G, Pallavicini P, Profumo A. 2016. A gold nanoparticle chemically modified gold electrode for the determination of surfactants. RSC Adv. 6:106500–106507.

Munawar H, Garcia-Cruz A, Majewska M, Karim K, Kutner W, Piletsky S. 2020. Electrochemical determination of fumonisin B1 using a chemosensor with a recognition unit comprising molecularly imprinted polymer nanoparticles. Sensors Actuators B Chem [Internet]. 321:128552. Available from: https://doi.org/10.1016/j.snb.2020.128552

Munawar H, Mankar JS, Sharma MD, Garcia-Cruz A, Fernandes LAL, Peacock M, Krupadam RJ. 2020. Highly selective electrochemical nanofilm sensor for detection of carcinogenic PAHs in environmental samples. Talanta [Internet]. 219:121273. Available from: https://doi.org/10.1016/j.talanta.2020.121273

Oberhaus F V., Frense D, Beckmann D. 2020. Immobilization Techniques for Aptamers on Gold Electrodes for the Electrochemical Detection of Proteins: A Review. Biosensors. 10.

Pagán M, Suazo D, del Toro N, Griebenow K. 2015. A comparative study of different protein immobilization methods for the construction of an efficient nano-structured lactate oxidase-SWCNT-biosensor. Biosens Bioelectron. 64:138–146.

Pereira S V., Bertolino FA, Fernández-Baldo MA, Messina GA, Salinas E, Sanz MI, Raba J. 2011. A microfluidic device based on a screen-printed carbon electrode with electrodeposited gold nanoparticles for the detection of IgG anti-Trypanosoma cruzi antibodies. Analyst. 136:4745–4751.

Ribone ME, Belluzo MS, Pagani D, Marcipar IS, Lagier CM. 2006. Amperometric bioelectrode for specific human immunoglobulin G determination: Optimization of the method to diagnose American trypanosomiasis. Anal Biochem. 350:61–70.

Saleh Ahammad AJ, Choi YH, Koh K, Kim JH, Lee JJ, Lee M. 2011. Electrochemical detection of cardiac biomarker troponin I at gold nanoparticle-modified ITO electrode by using open circuit potential. Int J Electrochem Sci. 6:1906–1916.

Sharma PS, Garcia-Cruz A, Cieplak M, Noworyta KR, Kutner W. 2019. ‘Gate effect’ in molecularly imprinted polymers: the current state of understanding. Curr Opin Electrochem [Internet]. 16:50–56. Available from: https://doi.org/10.1016/j.coelec.2019.04.020

Teles F, Tavira L, Fonseca L. 2010. Biosensors as rapid diagnostic tests for tropical diseases. . Cri Rev Cli Lab Sci. 47:139–169.

Theint H, Walsh J, Wong S, Voon K, Shitan M. 2019. Development of an optical biosensor for the detection of Trypanosoma evansi and Plasmodium berghei. Spectrochim Acta Part A Mol Biomol Spec. 218:348–358.

Wardhana A, Sawitri D. 2018. Surra: Trypanosomiasis pada ternak yang berpotensi sebagai penyakit zoonosis. Wartazoa. 28:139–151.

Wardhana A, Sawitri D. 2019. Deteksi Surra yang Disebabkan Trypanosoma evansi pada Kerbau di Kabupaten Pandeglang, Propinsi Banten. In: Martindah E, Wina E, editors. Semin Nas Teknol Peternak dan Vet “Mendukung Kemandirian Pangan di Era Ind 40.” Bogor: Pusat Penelitian dan Pengembangan Peternakan,Badan Penelitian dan Pengembangan Pertanian, Kementrian Pertanian; p. 451–461.

Welch NG, Scoble JA, Muir BW, Pigram PJ. 2017. Orientation and characterization of immobilized antibodies for improved immunoassays (Review). Biointerphases. 12:02D301.

Ying GQ, Wang MJ, Yi Y, Chen JS, Mei JF, Zhang YL, Chen SQ. 2018. Construction and application of an electrochemical biosensor based on an endotoxin aptamer. Biotechnol Appl Biochem. 65:323–327.

DOI: https://dx.doi.org/10.14334/Pros.Semnas.TPV-2020-p.844-857

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