DETEKSI GEN PENYANDI CLASSICAL STAPHYLOCOCCAL ENTEROTOXIN PADA Staphylococcus aureus ISOLAT ASAL HEWAN DAN MANUSIA

DHEAVY KUSUMA W.P.P, Prof. Dr. drh. Siti Isrina Oktavia Salasia

2023 | Skripsi | S1 KEDOKTERAN HEWAN

Angka kasus keracunan pangan di dunia selalu meningkat, penyebabnya adalah Staphylococcal enterotoxin yang dihasilkan oleh Staphylococcus aureus. Penelitian ini bertujuan untuk mendeteksi keberadaan gen penyandi classical enterotoxin Staphylococcus aureus pada isolat hewan dan manusia menggunakan teknik multiplex Polymerase Chain Reaction (PCR). Sebanyak 10 S. aureus isolat asal hewan yang digunakan dalam penelitian ini berasal dari Laboratorium Patologi Klinik FKH UGM, dan 10 S. aureus isolat asal manusia diperoleh dari Laboratorium Patologi Klinik FK UGM. Semua isolat diidentifikasi secara genotip dengan PCR. Gen enterotoksin S.aureus kemudian dianalisis menggunakan teknik multiplex PCR. Larutan 25 ul untuk PCR terdiri dari 2 ul primer forward (10pmol/ul), 2 ul primer reverse (10 pmol/ul), 12,5 ul PCR mix, 6,5 ul ddH2O dan 2 ul 20-40 ng DNA template. Program thermal cycler digunakan sesuai dengan referensi untuk setiap gen yang diamplifikasi. Hasil amplifikasi dianalisis menggunakan elektroforesis dengan agarose 2% dan Red Safe, kemudian divisualisasikan dengan transiluminator UV dibandingkan dengan kontrol dan penanda 1 Kb DNA Ladder. Semua isolat hewan dan manusia secara genotip positif untuk gen 23S rRNA, menunjukkan isolat S. aureus. Identifikasi gen enterotoksin menunjukkan bahwa S. aureus isolat asal manusia mengandung gen enterotoksin sec (80%), seh (60%), dan seg (20%). 10 S. aureus isolat asal hewan mendeteksi gen seh (100%), dan sec (80%). 10 S. aureus isolat asal manusia yang diisolasi mengandung kombinasi gen se(c,h) 50%, se(c,g)10%, se(c,g,h)10% dan pada isolat asal hewan gen se(c,h) 80%. Pada penelitian ini, gen enterotoksin yang paling sering terdeteksi adalah gen sec dan seh.

The number of cases of food poisoning in the world is continously increasing. Staphylococcal enterotoxin is the cause of food poisoning in humans and animals. One of the bacteria that produces enterotoxin is Staphylococcus aureus. This study aimed to detect the presence of the gene encoding the classical enterotoxin Staphylococcus aureus in animal and human isolates using the multiplex Polymerase Chain Reaction (PCR) technique. A total of 10 S. aureus animal isolates used in this study were derived from the Clinical Pathology Laboratory of Faculty of Veterinary Medicine UGM, and 10 S. aureus human isolates were obtained from the Clinical Pathology Laboratory of Faculty of medicine UGM. All isolates were genotypically identified by PCR. The enterotoxin gene of S.aureus were then analysed using a multiplex PCR technique. The 25 ul solution for PCR consist of 2 ul forward primer (10pmol/ul), 2 ul reverse primer (10 pmol/ul), 12,5 ul PCR mix, 6,5 ul ddH2O and 2 ul 20-40 ng DNA template. The thermal cycler program were used according to the references for each amplified genes. The amplification result were analyzed using electrophoresis with 2% agarose and Red Safe, and then visualized with a UV transilluminator compared to the control and 1 Kb DNA Ladder marker. All animal and human isolates were genotypically positive for 23S rRNA gene, indicating S. aureus isolates. The identification of enterotoxin genes showed that S. aureus human isolates contained enterotoxin sec (80%), seh (60%), and seg (20%) genes. Staphylococcus aureus animal isolates detected the seh (100%), and sec (80%) genes. Staphylococcus aureus human isolated contained a combination of genes of se(c,h) 50%, se(c,g)10%, se(c,g,h)10% and in animal isolates the gene se(c,h) 80%. In this study, the most frequently detected enterotoxin genes were the sec and seh genes.

Kata Kunci : Staphylococcus aureus, classical enterotoxin, gen, multiplex PCR