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All Journal HAYATI Journal of Biosciences Jurnal Veteriner dan Biomedis
Nugraha, Arifin
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Earth & Planetary Sciences Immunology & microbiology Medicine & Pharmacology Veterinary

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Development of Multiplex PCR for Simultaneous Detection of Trypanosoma evansi and Equine Piroplasma Infection in Horses Nugraha, Arifin; Cahyaningsih, Umi
HAYATI Journal of Biosciences Vol. 32 No. 3 (2025): May 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.3.740-746

Abstract

The identification of equine piroplasmosis and surra disease in infected horses currently presents a significant challenge. Clinical symptoms and blood smears are commonly used to diagnose these two diseases. The objective of this study was to develop a multiplex amplification assay capable of simultaneously identifying all three blood protozoa (equine piroplasma and Trypanosoma evansi) in a single test. The primer pairs used for detecting T. equi, B. caballi, and Trypanosoma evansi were ema-2-t (587 bp), Bc-134 (429 bp), and Rotat 1.2 VSG (151 bp), respectively. The multiplex PCR assay was subsequently evaluated for its detection limit, sensitivity percentage, and specificity using single PCR as the reference standard. The multiplex PCR method demonstrated a sensitivity of 100% for detecting both Theileria equi and Trypanosoma evansi, with all positive samples confirmed by a single PCR. It also achieved 100% specificity for both the parasites. However, for Babesia caballi, while the sensitivity remained at 100%, the specificity was reduced to 66%, indicating some limitations in accurately identifying negative samples. For the multiplex PCR assay, the minimum detectable concentrations were 0.01 ng/µL for both T. equi and B. caballi, whereas Trypanosoma evansi exhibited a detection threshold of 1 ng/µL. When multiplex PCR assays were used to screen blood samples obtained from horses in selected districts of Bogor, only a single sample showed a positive result for T. equi. The sensitivities and specificities achieved by multiplex PCR and single PCR were comparable. Therefore, the multiplex PCR method developed in this study can be effectively applied to diagnose piroplasmosis and surra in horses.
Pemberian Ekstrak Air Bidara Laut (Strychnos Ligustrina) terhadap Diferensial Leukosit Mencit yang diinfeksi dengan Plasmodium berghei Nurridho Wahid, Muhammad; Cahyaningsih, Umi; Nugraha, Arifin
Jurnal Veteriner dan Biomedis Vol. 2 No. 1 (2024): Maret
Publisher : Sekolah Kedokteran Hewan dan Biomedis

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jvetbiomed.2.1.35-42.

Abstract

Plasmodium berghei merupakan hemoprotozoa penyebab penyakit malaria pada mencit. Tujuan penelitian ini untuk mengetahui pengaruh ekstrak air bidara laut terhadap gambaran diferensial leukosit pada mencit yang diinfeksi Plasmodium berghei. Penelitian ini menggunakan 49 ekor mencit Balb/c, dibagi menjadi 7 kelompok yaitu kontrol normal (KN), kontrol infeksi tanpa perlakuan (KI), kontrol infeksi yang diberikan perlakuan obat kombinasi dihydroartemisinin 25 mg/kgbb dan piperakuin fosfat dosis 197 mg/kgbb (K0), EAa (ekstrak aquades), EAb (ekstrak aquades : DHF = 1:1), EAc (ekstrak aquades : DHF = 1:2), EAd (ekstrak aquades : DHF = 2:1). Kecuali KN, semua kelompok mencit diinfeksi Plasmodium berghei sebanyak 1 x 10 6 /ml secara intraperitoneal. Setiap hari diambil darah dari ekor mencit, dibuat apusan tipis dan dilakukan pewarnaan dengan Giemsa 10%, kemudian dilakukan perhitungan diferensial leukosit. Nilai rata-rata persentase diferensial leukosit diolah menggunakan uji ANOVA dan perbedaan hasil persentase pada masing-masing kelompok diketahui dengan menggunakan uji Duncan menggunakan software SPSS. Hasil penelitian menunjukkan bahwa perlakuan pada kelompok EAa dan EAd mengalami penurunan persentase neutrofil dan meningkatkan persentase limfosit pada hari ke 6 setelah infeksi. Persentase monosit, eosinofil dan basofil tidak mengalami perubahan yang signifikan. Pengobatan ekstrak air bidara laut dan DHF berpotensi sebagai obat antimalaria dengan menurunkan tingkat parasitemia dan meningkatkan persentase limfosit.
Co-Authors Nurridho Wahid, Muhammad Umi Cahyaningsih