AMPLIFIKASI GEN PENYANDI FUSION PROTEIN (F) VIRUS NEWCASTLE DISEASE (ND) DARI SPESIMEN LAPANGAN DENGAN METODE DIAGNOSA ONESTEP SIMPLEX RT-PCR

David Kristiawan, Dr. drh. Aris Haryanto, M.Si.

2011 | Skripsi | S1 KEDOKTERAN HEWAN

Virus Newcastle Disease(ND) termasuk ordo Mononegaviralesfamili Paramyxoviridaedengan genom ssRNA. Famili Paramyxoviridaememiliki 9 serotipe Avian paramyxovirus yaitu APMV-1 sampai APMV-9 berdasarkan uji serologis. Virus ND mempunyai dua protein yang terdapat pada bagian envelope, yaitu protein hemaglutinin/neuraminidasedan protein fusion. Pada bagian inti virus terdapat phosphoprotein, matrix protein, large protein, dan accessory proteins. Teknik diagnosa yang digunakan dalam penelitian ini adalah OneStep Simplex Reverse Transkriptase-Polymerase Chain Reaction (RT-PCR). Teknik tersebut mengamplifikasikan gen fusion virus ND dari spesimen lapangan tanpa adanya propagasi dan inokulasi virus dalam telur ayam berembrio (TAB). Penelitian ini menggunakan 8sampel swab tracheayang berasal dari BBVet Wates yang dikoleksi oleh drh. Loka Setia dari pasar Terban dan 7 sampel swab trachea dari BBVet Maros. Materi yang digunakan adalah Qiagen One Step RT-PCR Kit, QIAamp Viral RNA Kit, dan media elektroforesis menggunakan gel agarose 1,5 % dan SYBR Safe sebagai pewarna DNA agar terlihat di UV-transilluminator. Hasil penelitian menunjukkan bahwa dari 15 isolat yang digunakan 12 isolat positif virus ND dan 3 isolat negatif virus ND.

Newcastle Disease (ND) virus is included in the order Mononegavirales, family Paramyxoviridae with ssRNA genome. The family of Paramyxoviridae has nine serotypes of Avian paramyxovirus, APMV-1 until APMV-9 based from serologic test. ND virus has two proteins on the envelope, they are hemagglutinin/neuraminidase protein and fusion protein. At the core of the virus there are phosphoprotein, matrix protein, large protein, and accessory proteins. The Diagnosa technique being used in this research is OneStep Simplex Reverse Transkriptase-Polymerase Chain Reaction (RTPCR). This technique amplified fusion gene of ND virus from field specimens without propagation and virus inoculation in embryonated chicken eggs. This study used eight swab trachea samples from BBVet Wates which were collected by drh. Loka Setia from Terban market and seven swab trachea samples from BBVet Maros. The material are Qiagen One Step RT-PCR Kit, QIAamp Viral RNA Kit and the media electrophoresis using 1.5% agarose gel and SYBR Safe as a colorant DNA to be visible in the UV-transilluminator. The result was from the 15 isolates, 12 of them have positive reaction to ND virus and 3 isolates have negative reaction to ND virus.

Kata Kunci : Newcastle Disease,protein, hemaglutinin/neuramidase, fusion, RT-PCR