Abstract
Numerous viral pathogens are circulated in the environment of commercial chicken farms that causing the difficulty in the confirmation of diagnosis. A breakthrough on the diagnosis technique is required in order to deal with multiple viral infections. Ideally, the approach should have not only high accuracy but also economical and straightforward. The objective of this research is to develop a rapid multiplex reverse transcriptase polymerase chain reaction (mRT-PCR) diagnostic method for three common infectious viral diseases in poultry, Newcastle Disease (ND), Avian Influenza (AI) subtype H5N1 and Infectious Bronchitis (IB). This study was successful in developing and optimizing the mRT-PCR for these three pathogens in a single reaction test. Testing 67 field samples from Sukabumi district revealed the presence of several targeted viruses. Key Words : Screening Test, Multiplex RT-PCR, Newcastle Disease, Avian Influenza, Infectious Bronchitis
Highlights
Virus standar dan sampel lapanganVirus standard yang dipergunakan sebagai kontrol positif dalam tahapan optimasi uji multiplex reverse transcriptase polymerase chain reaction (mRT-PCR) berasal dari isolat koleksi laboratorium Virologi BBalitvet dan strain vaksin dari sumber komersial
Numerous viral pathogens are circulated in the environment of commercial chicken farms that causing the difficulty in the confirmation of diagnosis
A breakthrough on the diagnosis technique is required in order to deal with multiple viral infections
Summary
Virus standard yang dipergunakan sebagai kontrol positif dalam tahapan optimasi uji mRT-PCR berasal dari isolat koleksi laboratorium Virologi BBalitvet dan strain vaksin dari sumber komersial. Kontrol positif untuk virus ND digunakan isolat ITA dan RIVS. Untuk virus IB digunakan isolat PTS3 yang merupakan koleksi. Sampel lapangan (swab kloaka dan organ) yang diperlukan untuk menguji protokol mRT-PCR dikoleksi dari beberapa peternakan komersial di Kabupaten. Sampel dijaga pada suhu 4oC dan dibawa ke laboratorium virologi BBalitvet untuk disimpan pada. Primer spesifik untuk uji RT-PCR dalam mendeteksi virus ND, AI dan IB ditentukan berdasarkan marker gen yang akan diamplifikasi sesuai dengan beberapa penelitian terdahulu (Stäuber et al 1995; Jackwood et al 1997; Gohm et al 2000; Lee et al 2000; Lee et al 2001; Dharmayanti et al 2004; Meir et al 2010).
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