Karakterisasi hemolisin dan analisis amplified fragment length polymorphism (AFLP) Staphylococcus aureus isolat pangan olahan asal hewan

ARIYANTI, Dwi, Prof. Dr. drh. Siti Isrina Oktavia Salasia

2010 | Tesis | S2 Bioteknologi

Staphylococcus aureus merupakan patogen penting pada manusia yang sering menimbulkan kasus penyakit dan keracunan pangan. Patogenesitas dan virulensi Staphylococcus aureus ditentukan oleh substansi yang diproduksinya, antara lain toksin dan enzim perusak. Hemolisin merupakan salah satu faktor virulen Staphylococcus aureus yang diproduksi oleh hampir seluruh strain. Penelitian ini bertujuan untuk melakukan karakterisasi hemolisin secara fenotipik dan genotipik, serta analisis hubungan klonal antar isolat Staphylococcus aureus yang berasal dari pangan olahan asal hewan. Dalam penelitian ini digunakan 11 Staphylococcus aureus isolat dari berbagai pangan olahan asal hewan yang diambil dari pasar tradisional dan supermarket di Yogyakarta, Sidoarjo, Jakarta dan Bandung. Karakterisasi hemolisin secara fenotipik dilakukan berdasar pengamatan sifat hemolisis dengan Staphylococcus aureus pada plat agar darah domba. Karakterisasi genotipik dilakukan dengan mengamplifikasi gen penyandi hemolisin dengan teknik polymerase chain reaction (PCR) menggunakan primer spesifik. Hubungan klonal antar isolat Staphylococcus aureus dianalisis menggunakan amplified fragment length polymorphism (AFLP). Secara fenotipik Staphylococcus aureus memperlihatkan sifat hemolisis alfa (18,18%), hemolisis beta (27,27%) dan hemolisis gama (54,55%) pada plat agar darah domba. Hasil karakterisasi hemolisin secara genotipik diketahui bahwa Staphylococcus aureus mengandung gen hla (81,81%), kombinasi gen hlb dan hla (18,18%). Hasil analisis AFLP berdasar perhitungan unweight pair group method with arithmatic average (UPGMA) menunjukkan bahwa Staphylococcus aureus dari berbagai pangan olahan asal hewan dapat dikelompokkan menjadi 5 klaster. Hasil penelitian ini diharapkan dapat digunakan sebagai dasar dalam menentukan strategi kontrol stafilokokosis.

Staphylococcus aureus is an important pathogen of animal and human which caused many cases diseases and food poisoning. Virulence and pathogenecity of Staphylococcus aureus could be determined by the production of toxins and enzymes. Haemolysin is one of the virulence factor of Staphylococcus aureus that produced by almost strains. The aims of the research were to characterize haemolysin of Staphylococcus aureus phenotypic and genotypically, and to analyse clonal association among isolates of Staphylococcus aureus from various food of animal origins. The research were used 11 Staphylococcus aureus isolated from various food of animal origins from traditional markets and supermarkets in Yogyakarta, Sidoarjo, Jakarta and Bandung. Characterization of haemolysin phenotypically based on haemolysis pattern of Staphylococcus aureus on sheep blood agar plate. Gene encoding hemolysin were amplified with specific primers by using polymerase chain reaction (PCR) technique. Clonal relationship among Staphylococcus aureus isolates were analysed by amplified fragment length polymorphism (AFLP). The results of the studies showed that Staphylococcus aureus on sheep blood agar plates revealed an alpha haemolysis pattern (18,18%), beta haemolysis (27,27%) and gamma haemolysis (54,55%). Based on amplification of the gene encoding haemolysin of Staphylococcus aureus with specific primers showed hla genes (81,81%), and hla combined with hlb genes (18,18%). The size of amplicon for hla gene was approximately 534 bp and 833 bp for hlb gene. The results of the AFLP analysis with unweight pair group method with arithmatic average (UPGMA) of Staphylococcus aureus isolated from food from animal origins could be grouped into 5 clusters. The mapping of Staphylococcus aureus based on characters of haemolysin and genetic relationship might help to control staphylococcosis.

Kata Kunci : Staphylococcus aureus,Hemolisin,PCR,AFLP,Pangan olahan asal hewan,Staphylococcus aureus, haemolisyn, PCR, AFLP, food of animal origin