Single-Step Multiplex Reverse Transcription–Polymerase Chain Reaction (RT-PCR) for Influenza A Virus Subtype H5N1 Detection

We analyzed the data available in Nepal during this pandemic in order to determine the epidemiological, clinical and virological characteristics of pandemic influenza A in 2009. The test was conducted by real-time Reverse Transcription – Polymerase Chain Reaction on sample from patients with suspected influenza-like illnesses. Out of 538 cases were tested, 32 % were positive for pandemic influenza A 2009 and the infection rate was highest for cases of 11-20 years and lowest in >50 years of age. Keywords: Influenza A ; pandemic; RT-PCR; surveillance.

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The Single-Step Multiplex Reverse Transcription- Polymerase Chain Reaction Assay for Detecting H5 and H7 Avian Influenza A Viruses

The Tohoku Journal of Experimental Medicine ◽

10.1620/tjem.211.75 ◽

2007 ◽

Vol 211 (1) ◽

pp. 75-79 ◽

Cited By ~ 9

Author(s):

Aunyaratana Thontiravong ◽

Sunchai Payungporn ◽

Juthatip Keawcharoen ◽

Salin Chutinimitkul ◽

Sumitra Wattanodorn ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Avian Influenza ◽

Reverse Transcription ◽

Polymerase Chain Reaction Assay ◽

Influenza A ◽

Single Step ◽

Influenza A Viruses ◽

Chain Reaction ◽

Polymerase Chain

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PENENTUAN SUBTIPE VIRUS AVIAN INFLUENZA DENGAN METODE SINGLE STEP MULTIPLEX REVERSE TRANSCRIPTASE- POLYMERASE CHAIN REACTION (RT-PCR) ISOLAT ASAL PROVINSI ACEH

Jurnal Kedokteran Hewan - Indonesian Journal of Veterinary Sciences ◽

10.21157/j.ked.hewan.v8i1.1265 ◽

2014 ◽

Vol 8 (1) ◽

Author(s):

Teuku Zahrial Helmi ◽

Rini Widayanti ◽

Aris Haryanto

Keyword(s):

Polymerase Chain Reaction ◽

Avian Influenza ◽

Reverse Transcriptase ◽

Influenza A ◽

Deoxyribonucleic Acid ◽

Single Step ◽

Rt Pcr ◽

Chain Reaction ◽

Polymerase Chain ◽

Virus Influenza

Tujuan dari penelitian ini adalah mengidentifikasi keberadaan gen M, H5, dan N1 virus avian influenza (AI) melalui metode single step multiplex reverse transcriptase-polymerase chain reaction (RT-PCR), sebagai acuan untuk peneguhan diagnosis secara molekuler virus AI di Provinsi Aceh. Penelitian ini menggunakan 11 isolat virus AI asal Provinsi Aceh yang diperoleh dari Laboratorium Balai Penyidikan dan Pengujian Veteriner (BPPV) Regional I Medan di Sumatera Utara dari tahun 2006-2008. Penelitian dilakukan di Laboratorium Virologi BPPV Regional I di Medan, Sumatera Utara. Amplifikasi terhadap gen matriks (M) virus AI menggunakan metode simplex RT-PCR. Hasil simplex RT-PCR terhadap gen M diperoleh 10 isolat yang menunjukkan pita deoxyribonucleic acid (DNA) pada 276 bp dan satu isolat yang tidak muncul, kemudian dilanjutkan dengan metode single step multiplex RT-PCR menggunakan pasangan primer gen penyandi protein N1, H5, dan M. Produk PCR 131 bp (N1), 189 bp (H5), dan 276 bp (M) muncul sebagai hasil elektroforesis dari semua isolat virus AI. Semua virus AI yang mewabah dari tahun 2006-2008 di Provinsi Aceh termasuk ke dalam virus influenza A subtipe H5N1.

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Detection of Avian Influenza Virus of H9 Subtype in the Faeces of Experimentally and Naturally Infected Chickens by Reverse Transcription-Polymerase Chain Reaction

Acta Veterinaria Brno ◽

10.2754/avb200776030405 ◽

2007 ◽

Vol 76 (3) ◽

pp. 405-413 ◽

Cited By ~ 2

Author(s):

H. Noroozian ◽

M. Vasfi Marandi ◽

M. Razazian

Keyword(s):

Polymerase Chain Reaction ◽

Avian Influenza ◽

Reverse Transcription ◽

Predictive Value ◽

Relative Sensitivity ◽

Rt Pcr ◽

Chain Reaction ◽

Diagnostic Laboratory ◽

Polymerase Chain ◽

Virus Subtype

Avian Influenza (AI) is a viral, highly contagious disease of domestic and wild birds. In an avian diagnostic laboratory, it is essential to have methods for rapid detection of respiratory viruses. In the present study, cloacal swabs collected from chickens experimentally and naturally infected with mild pathogenicity AI virus subtype H9, used in a reverse transcription-polymerase chain reaction (RTPCR) assay for detection of AI. On cloacal swabs collected from experimentally infected chickens, AI virus was detected most frequently between days 3 and 7 post infection (p.i.) and the relative sensitivity, specificity, correlation rate, positive predictive value and negative predictive value of the RT-PCR compared to virus isolation (VI) assay were 84%, 80%, 82%, 83% and 81%, respectively. On pooled cloacal swabs collected from flocks suspected of AI, these results were 96%, 100%, 97%, 83% and 100%, respectively. The results proved that the RT-PCR assay could be a reliable and rapid alternative to VI assay for detection of AI virus subtype H9 in faecal specimens.

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