Deteksi Molekuler Plasmodium sp. pada Macaca fascicularis Menggunakan Penanda Gen Sitokrom b dan SSU rRNA dari Sampel Darah dan Usap Rektal

Abstract

Plasmodium sp. merupakan parasit penyebab penyakit malaria pada manusia. Dalam siklus penularannya, monyet ekor panjang (Macaca fascicularis) berperan sebagai inang penting penyakit zoonosis ini. Informasi mengenai deteksi infeksi Plasmodium sp. pada primata masih terbatas, terutama pemilihan penanda gen molekuler dan penggunaan sampel non-invasif. Penelitian ini bertujuan mendeteksi Plasmodium sp. pada M. fascicularis menggunakan penanda gen sitokrom b (cyt b) dan small subunit ribosomal RNA (SSU rRNA), membandingkan dan menghitung persentase keberhasilan amplifikasi DNA sampel darah dan usap rektal. Deteksi Plasmodium terhadap 23 sampel darah dan 23 sampel usap rektal dilakukan menggunakan PCR dengan marka gen sitokrom b (170 pb dan 90 pb) dan gen SSU rRNA parsial (1640 pb) lalu divisualisasi menggunakan elektroforesis dengan gel agarose 2% dan pewarna ethidium bromida. Seluruh sampel darah terdeteksi positif Plasmodium sp. pada kedua gen penanda sedangkan sampel usap rektal hanya menghasilkan amplikon pada sitokrom b (90 pb) sebesar 13% yaitu W0163, W0139 dan 170702A. Hasil-hasil ini mengindikasikan dapat dilakukannya deteksi Plasmodium sp. melalui sampel usap rektal, meskipun tingkat keberhasilannya lebih kecil dibandingkan sampel darah pada M. fascicularis.

Plasmodium sp. is a major parasitic protozoan responsible for malaria in humans. In its transmission cycle, long-tailed macaques (Macaca fascicularis) serve as important reservoir hosts. However, information of detecting malaria infections in non-human primates remains limited. In this study, Plasmodium sp. infection in M. fascicularis was detected using cytochrome b (cyt b) and small subunit ribosomal RNA (SSU rRNA) gene markers, followed by calculation and comparison of DNA amplification success rates between blood and rectal swab samples. DNA was isolated from 23 blood samples and 23 rectal swab samples. Detection was performed using PCR targeting cytochrome b fragments (170 bp and 90 bp) and a partial SSU rRNA gene fragment (1640 bp), with amplification products visualized by 2% agarose gel electrophoresis stained with ethidium bromide. All blood samples tested positive for Plasmodium sp. for both gene markers. In contrast, detection from rectal swab samples was successful in only 13% of samples using the cytochrome b 90 bp marker, specifically in samples W0163, W0139, and 170702A.These findings demonstrate that Plasmodium sp. can be detected from rectal swab samples; however, the detection success rate is substantially lower than that obtained from blood samples in M. fascicularis.